Estrogen receptor ligands and methods of use thereof

ABSTRACT

The present invention relates to methods for reducing testosterone levels by reduction of luteinizing hormone (LH) or independent of LH levels in a male subject and methods of treating, suppressing, reducing the incidence, reducing the severity, or inhibiting prostate cancer, advanced prostate cancer, and castration-resistant prostate cancer (CRPC) and palliative treatment of prostate cancer, advanced prostate cancer and castration-resistant prostate cancer (CRPC). The compounds of this invention suppress free or total testosterone levels to castrate levels which may be used to treat prostate cancer, advanced prostate cancer, and CRPC without causing bone loss, decreased bone mineral density, increased risk of bone fractures, increased body fat, hot flashes and/or gynecomastia.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a Continuation in Part of International ApplicationNumber PCT/US10/25032, filed Feb. 23, 2010 which claims priority to U.S.Ser. No. 61/154,707, filed Feb. 23, 2009; and U.S. Ser. No. 61/168,983filed Apr. 14, 2009; and United-States Ser. No. 61/261,669, filed Nov.16, 2009; and claims priority to U.S. Ser. No. 61/380,113, filed Sep. 3,2010, each of which is hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to methods for reducing testosteronelevels by reduction of luteinizing hormone (LH) or independent of LHlevels in a male subject and methods of treating, suppressing, reducingthe incidence, reducing the severity, or inhibiting advanced prostatecancer and castration-resistant prostate cancer (CRPC) and palliativetreatment of advanced prostate cancer and castration-resistant prostatecancer (CRPC).

BACKGROUND OF THE INVENTION

Estrogens refer to a group of endogenous and synthetic hormones that areimportant for and used for tissue and bone maintenance. Estrogens areendocrine regulators in the cellular processes involved in thedevelopment and maintenance of the reproductive system. The role ofestrogens in reproductive biology, the prevention of postmenopausal hotflashes, and the prevention of postmenopausal osteoporosis are wellestablished. Estradiol is the principal endogenous human estrogen, andis found in both women and men.

The biological actions of estrogens and antiestrogens are manifestthrough two distinct intracellular receptors, estrogen receptor alpha(ERα) and estrogen receptor beta (ERβ). Endogenous estrogens aretypically potent activators of both receptor subtypes. For exampleestradiol acts as an ERα agonist in many tissues, including breast,bone, cardiovascular and central nervous system tissues. Selectiveestrogen receptor modulators commonly act differently in differenttissues. For example, a SERM may be an ERα antagonist in the breast, butmay be a partial ERα agonist in the uterus, bone and cardiovascularsystems. Compounds that act as estrogen receptor ligands are, therefore,useful in treating a variety of conditions and disorders.

Prostate cancer is one of the most frequently diagnosed noncutaneouscancers among men in the US and is the second most common cause ofcancer deaths with over 180,000 new cases and almost 29,000 deathsexpected this year. Patients with advanced prostate cancer undergoandrogen deprivation therapy (ADT), typically either by luteinizinghormone releasing hormone (LHRH) agonists or by bilateral orchiectomy.Androgen deprivation therapy not only reduces testosterone, but estrogenlevels are also lower since estrogen is derived from the aromatizationof testosterone, which levels are depleted by ADT. Androgen deprivationtherapy-induced estrogen deficiency causes significant side effectswhich include hot flushes, gynecomastia and mastalgia, bone loss,decreases in bone quality and strength, osteoporosis andlife-threatening fractures, adverse lipid changes and highercardiovascular disease and myocardial infarction, and depression andother mood changes. It is believed that many of the estrogen deficiencyside effects of ADT are mediated by ERα.

Leuprolide acetate (Lupron®) is a synthetic nonapeptide analog ofnaturally occurring gonadotropin-releasing hormone (GnRH or LHRH).Leuprolide acetate is an LHRH superagonist that eventually suppresses LHsecretion by the pituitary. Leuprolide acetate acts as a potentinhibitor of gonadotropin secretion, resulting in suppression of ovarianand testicular steroidogenesis. In humans, administration of leuprolideacetate results in an initial increase in circulating levels ofluteinizing hormone (LH) and follicle stimulating hormone (FSH), leadingto a transient increase in levels of the gonadal steroids (testosteroneand dihydrotestosterone in males, and estrone and estradiol inpremenopausal females). However, continuous administration of leuprolideacetate results in decreased levels of LH and FSH. In males,testosterone is reduced to castrate levels (below 50 ng/dL). Inpremenopausal females, estrogens are reduced to postmenopausal levels.Testosterone is a known stimulus for cancerous cells of the prostate.Suppressing testosterone secretion or inhibiting the actions oftestosterone is thus a necessary component of prostate cancer therapy.Leuprolide acetate can be used for LH suppression, which is thereduction and lowering of serum testosterone to castrate levels to treatprostate cancer.

Prior to the introduction of LHRH agonists, castrate testosterone levelswere achieved by increasing estrogen activity in the pituitary viaestrogens, primarily diethylstilbestrol (DES). DES was equally effectiveas LHRH agonists at suppressing testosterone to castrate levels.Patients treated with DES did not have hot flashes or bone loss, but didhave gynecomastia at higher rates than ADT with LHRH agonists.Unfortunately, highly potent, pure estrogens, like DES and estradiol,are often associated with a high risk of severe cardiovascular andthromboembolic complications which have limited their clinical use.

The compounds of this invention suppress testosterone levels to castratelevels which may be used to treat prostate cancer, while preventing theincreased risk of thrombotic events, and without causing bone loss, hotflashes and/or gynecomastia.

The compounds of this invention suppress free or total testosteronelevels to castrate levels which may be used to treat prostate cancer,advanced prostate cancer, or castration-resistant prostate cancerwithout causing bone loss, decreased bone mineral density, increasedrisk of bone fractures, increased body fat, hot flashes, and/orgynecomastia.

SUMMARY OF THE INVENTION

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound of this invention asdescribed herein below.

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound of this invention asdescribed herein below, wherein the lowering of total serum testosteroneoccurs by a reduction of serum luteinizing hormone levels.

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound of this invention asdescribed herein below, wherein the lowering of total serum testosteroneis independent of a reduction of serum luteinizing hormone levels.

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound of this invention asdescribed herein below, wherein said administering said compoundprevents or treats side effects associated with androgen deprivationtherapy (ADT) from occurring, wherein said subject has prostate cancer.

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound of this invention asdescribed herein below, wherein administering the compound produces alowering of total serum testosterone without causing side effectsassociated with androgen deprivation therapy (ADT).

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound of this invention asdescribed herein below, wherein administering the compound produces alowering of total serum testosterone without causing hot flashes,gynecomastia, increased body fat, decreased bone mineral density, orincreased risk of bone fracture.

In one embodiment, this invention provides a method for androgendeprivation therapy in a subject comprising administering atherapeutically effective amount of a compound of this invention asdescribed herein below. In another embodiment, said subject has prostatecancer.

In one embodiment this invention provides a method of treating,suppressing, reducing the incidence, reducing the severity, orinhibiting advanced prostate cancer comprising administering atherapeutically effective amount of a compound of this invention asdescribed herein below.

In one embodiment this invention provides a method of increasingsurvival of a subject with advanced prostate cancer comprisingadministering a therapeutically effective amount of a compound of thisinvention as described herein below.

In one embodiment this invention provides a method of palliativetreatment of advanced prostate cancer comprising administering atherapeutically effective amount of a compound of this invention asdescribed herein below.

A method of treating, suppressing, reducing the incidence, reducing theseverity, or inhibiting the progression of castration-resistant prostatecancer (CRPC) and its symptoms or increasing the survival of men withcastration-resistant prostate cancer comprising administering atherapeutically effective amount of a compound of this invention, asdescribed herein below.

In one embodiment, this invention provides a method of increasingsurvival of a subject with castration-resistant prostate cancer (CRPC)comprising administering a therapeutically effective amount of acompound of this invention as described herein below.

In one embodiment, this invention provides a method of palliativetreatment of castration-resistant prostate cancer (CRPC) comprisingadministering a therapeutically effective amount of a compound of thisinvention as described herein below.

In one embodiment, this invention is directed to a method of increasingserum concentrations of sex or steroid hormone binding globulin (SHBG).In one embodiment, this invention is directed to a method of decreasingserum, tissue or tumor concentrations of free or bioavailabletestosterone via increasing the SHBG comprising administering atherapeutically effective amount of this invention.

In one embodiment, the methods of this invention comprise administeringa combination of a compound of this invention with a LHRH agonist. Inanother embodiment, the LHRH agonist is Lupron®.

In one embodiment, the compounds of this invention suppress free ortotal testosterone levels to castrate levels which may be used to treatprostate cancer, advanced prostate cancer, or castration-resistantprostate cancer without causing bone loss, decreased bone mineraldensity, increased risk of bone fractures, increased body fat, hotflashes, and/or gynecomastia.

BRIEF DESCRIPTION OF THE DRAWINGS

The subject matter regarded as the invention is particularly pointed outand distinctly claimed in the concluding portion of the specification.The invention, however, both as to organization and method of operation,together with objects, features, and advantages thereof, may best beunderstood by reference to the following detailed description when readwith the accompanying drawings in which:

FIG. 1 depicts serum testosterone (solid line) and total androgen(dotted line) levels in intact male monkeys after daily 30 mg/kg oraladministration of Compound IV (first dose on Day 0). (See Example 8.)

FIG. 2 depicts testosterone levels in intact rats treated with CompoundIV (0.3, 1, 10, 30 mg/kg). ^(I)denotes P<0.05 vs. intact vehiclecontrols. BLOQ values are represented graphically at the limit ofquantitation 0.08 ng/mL. (See Example 9.)

FIG. 3 depicts the inhibitory effect of Compound IV on 17β-HSD5 enzymeactivity. (See Example 12.)

FIG. 4 depicts in vitro aggregation of human platelets in the presenceof DES, 17β-estradiol (E2), and Compound IV. Platelet Rich Plasma (PRP)was incubated with vehicle, E2, DES, or Compound IV for 30 secondsbefore inducing aggregation with 0.3 units of thrombin. Aggregation wasmonitored for 5 minutes and expressed as a percentage of vehiclecontrol. (See Example 13.)

FIG. 5 Generic synthetic scheme for the preparation of Compounds II-XII.(See Example 1.)

FIG. 6 Synthetic scheme for the preparation of Compound IV. (See Example2.)

FIG. 7 Synthetic scheme for the preparation of Compound VI. (See Example3.)

FIG. 8 Synthetic scheme for the preparation of Compounds I× and X. (SeeExample 5.)

FIG. 9 depicts testosterone levels in intact rats treated with CompoundIV after 24 h, 72 h and 168 h with dosages of 3 mg/kg, 10 mg/kg and 300mg/kg. (See Example 9)

FIG. 10 depicts LH levels (FIG. 10A), FSH levels (FIG. 10B),testosterone levels (FIG. 10C), prostate weight levels (FIG. 10D),seminal vesicle weight levels (FIG. 10E) and levator ani weight (FIG.10F) of treated intact and orchidectomized (ORX) rats with 0.3 mg/kg, 1mg/kg, 3 mg/kg, 10 mg/kg and 30 mg/kg dosages of Compound IV.^(I)denotes P<0.05 vs. intact vehicle controls. ^(O)denotes P<0.05 vs.ORX vehicle controls BLOQ values are represented graphically at thelimit of quantitation 0.08 ng/mL. (See Example 9.)

FIG. 11 depicts prostate size in intact and ORX rats by administeringCompound IV (FIG. 11A) and DES (FIG. 11B) at different dosages. (SeeExample 15.)

FIG. 12 depicts differences between DES and Compound IV; DES crossreactswith glucocorticoid receptor (GR) while Compound IV does not (FIG. 12A);DES crossreacts with androgen receptor (AR). It mildly stimulates ARaction and mildly inhibits (i.e., it is a partial agonist/antagonist)while Compound IV does not (FIG. 12B); DES abrogates estrogen relatedreceptor (ERR) transactivation, while Compound IV does not (FIG. 12C).(See Example 15.)

FIG. 13 depicts effect of Compound IV on attenuation of hot flashes inmorphine withdrawal model with 5 mg/kg, 10 mg/kg, 15 mg/kg and 30 mg/kgdosages. N=7 animals per group. 17β-E2 was used at 5 mg/kg in 100% DMSO.(See Example 14.)

FIG. 14 depicts dose dependent body weight (kg) reductions of monkeys(˜20% at 100 mg/kg) by administering Compound IV for 91 days. No sign ofgynecomastia or hyperestrogenicity was observed. (See Example 16.)

FIG. 15 depicts dose dependent serum testosterone level reductions(ng/mL) in monkeys after daily oral administration of Compound IVcompared to positive control (LHRH agonist). Dotted line indicates thetestosterone level of chemically castrated patients and the bold dashedline indicates the testosterone level of surgically castrated monkeys.(See Example 16.)

FIG. 16 depicts dose dependent prostate-specific antigen (PSA) levels(ng/mL) in monkeys by administering Compound IV at baseline and at day28. PSA levels were significantly decreased with Compound IV treatment.(See Example 16.)

FIG. 17 depicts dose dependent prostate volume using transrectalultrasound (TRUS) in monkeys compared to positive control (LHRHagonist), by administering Compound IV at week 6. (See Example 16.)

FIG. 18 depicts dose dependent organ weights (prostate, seminal vesicleand testis) as percent of control of monkeys at day 90, by administeringCompound IV (FIG. 18A). Prostate weights at 13-week necropsy in monkeysafter daily oral administration of Compound IV (FIG. 18B). (See Example16.)

FIG. 19 depicts dose dependent mean total testosterone levels (nmol/L)in humans for a period between 1-11 days by administering Compound IV(100 mg, 300 mg, 600 mg and 1000 mg). (See Example 17.)

FIG. 20 depicts dose dependent mean LH levels (IU/L) in humans for aperiod between 1-10 days by administering compound IV (100 mg, 300 mg,600 mg and 1000 mg. (See Example 17.)

FIG. 21 depicts dose dependent mean free testosterone levels (pg/mL) inhumans for a period between 1-10 days by administering compound IV (100mg, 300 mg, 600 mg and 1000 mg. (See Example 17.)

FIG. 22 depicts dose dependent mean PSA levels (μg/L) in humans for aperiod between 1-10 days by administering compound IV (100 mg, 300 mg,600 mg and 1000 mg). (See Example 17.)

FIG. 23 depicts dose dependent serum testosterone levels (ng/mL) inintact rats after 14 days recovery of administering Compound IV.^(I)denotes P<0.05 vs Intact controls. (See Example 10.)

It will be appreciated that for simplicity and clarity of illustration,elements shown in the figures have not necessarily been drawn to scale.For example, the dimensions of some of the elements may be exaggeratedrelative to other elements for clarity. Further, where consideredappropriate, reference numerals may be repeated among the figures toindicate corresponding or analogous elements.

DETAILED DESCRIPTION OF THE PRESENT INVENTION

In the following detailed description, numerous specific details are setforth in order to provide a thorough understanding of the invention.However, it will be understood by those skilled in the art that thepresent invention may be practiced without these specific details. Inother instances, well-known methods, procedures, and components have notbeen described in detail so as not to obscure the present invention.

In one embodiment, the compounds as described herein and/or compositionscomprising the same may be used for lowering total serum testosteronelevels in a male subject.

In one embodiment, the compounds as described herein and/or compositionscomprising the same may be used for lowering total serum testosteronelevels and lowering prostate specific antigen (PSA) in a male subject.In one embodiment, the lowering of total serum testosterone levels is tocastrate levels. In one embodiment, the lowering of total serumtestosterone levels does not reach castrate levels.

In one embodiment, the compounds as described herein and/or compositioncomprising the same may be used for lowering prostate specific antigen,independent of reduction or lack thereof on testosterone levels.

In one embodiment, the compounds as described herein and/or compositionscomprising the same may be used for lowering total serum testosteronelevels in a male subject wherein the lowering of total serumtestosterone occurs by a reduction of serum luteinizing hormone (LH)levels.

In one embodiment, the compounds as described herein and/or compositionscomprising the same may be used for lowering total serum testosteronelevels in a male subject wherein the lowering of total serumtestosterone is independent of a reduction of serum luteinizing hormonelevels.

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound or its isomer,pharmaceutical acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof, represented by the structure offormula I:

wherein

Y is C(O) or CH₂;

R₁, R₂ are independently hydrogen, halogen, hydroxyl, alkoxy, cyano,nitro, CF₃, N(R)₂, sulfonamide, SO₂R, alkyl, haloalkyl, aryl,O-Alk-NR₅R₆ or O-Alk-heterocycle in which the heterocycle is a 3-7membered substituted or unsubstituted heterocyclic ring, optionallyaromatic;

R₃, R₄ are independently hydrogen, halogen, hydroxyalkyl, hydroxyl,alkoxy, cyano, nitro, CF₃, NHCOR, N(R)₂, sulfonamide, SO₂R, alkyl,haloalkyl, aryl or protected hydroxyl;

R is alkyl, hydrogen, haloalkyl, dihaloalkyl, trihaloalkyl, CH₂F, CHF₂,CF₃, CF₂CF₃, aryl, phenyl, halogen, alkenyl, CN, NO₂, or OH;

R₅ and R₆ are independently hydrogen, phenyl, an alkyl group of 1 to 6carbon atoms, a 3 to 7 membered cycloalkyl, a 3 to 7 memberedheterocycle, a 5 to 7 membered aryl; or R₅ and R₆ form a 3 to 7 memberedring with the nitrogen atom;

j and k are independently 1-4; and

Alk is linear alkyl of 1-7 carbons, branched alkyl of 1-7 carbons, orcyclic alkyl of 3-8 carbons.

In additional embodiments of the methods described herein, the compoundof Formula I is represented by formula IA:

wherein R₁, R₂, R₃, R₄, j and k are as defined for Formula I.

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound or its isomer,pharmaceutical acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof, represented by a compound of formulaII:

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound or its isomer,pharmaceutical acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof, represented by a compound of formulaIII:

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound or its isomer,pharmaceutical product, pharmaceutical acceptable salt, polymorph,hydrate or any combination thereof, represented by a compound of formulaIV:

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound or its isomer,pharmaceutical acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof, represented by a compound of formulaV:

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound or its isomer,pharmaceutical acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof, represented by a compound of formulaVI:

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound or its isomer,pharmaceutical acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof, represented by a compound of formulaVII:

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound or its isomer,pharmaceutical acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof, represented by a compound of formulaVIII:

In one embodiment, this invention provides a method of lowering totalserum testosterone levels by reduction of luteinizing hormone (LH)levels in a male subject having prostate cancer, comprisingadministering a therapeutically effective amount of a compound or itsisomer, pharmaceutical acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof represented by a compoundof formula IX:

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound or its isomer,pharmaceutical acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof, represented by a compound of formulaX:

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound or its isomer,pharmaceutical acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof, represented by a compound of formulaXI:

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound or its isomer,pharmaceutical acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof, represented by a compound of formulaXII:

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound of formula IA, I-XII orits isomer, pharmaceutical acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof. In another embodiment themale subject has prostate cancer. In another embodiment, the total serumtestosterone is lowered below about 100 ng/dL. In another embodiment,the total serum testosterone is lowered below about 50 ng/dL. In anotherembodiment, the total serum testosterone concentration is lowered belowabout 25 ng/dL.

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound of formula IA, I-XII orits isomer, pharmaceutical acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof, wherein the lowering oftotal serum testosterone occurs by a reduction of serum luteinizinghormone (LH) levels. In another embodiment the male subject has prostatecancer. In another embodiment, the total serum testosterone is loweredbelow about 100 ng/dL. In another embodiment, the total serumtestosterone is lowered below about 50 ng/dL. In another embodiment, thetotal serum testosterone concentration is lowered below about 25 ng/dL.

In one embodiment, this invention provides a method of lowering freeserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound of formula IA, I-XII orits isomer, pharmaceutical acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof, wherein the lowering offree serum testosterone occurs by a reduction of serum luteinizinghormone (LH) levels. In another embodiment the male subject has prostatecancer.

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound of formula IA, I-XII orits isomer, pharmaceutical acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof, wherein the lowering oftotal serum testosterone is independent of a reduction of serumluteinizing hormone (LH) levels. In another embodiment the male subjecthas prostate cancer. In another embodiment, the total serum testosteroneis lowered below about 100 ng/dL. In another embodiment, the total serumtestosterone is lowered below about 50 ng/dL. In another embodiment, thetotal serum testosterone concentration is lowered below about 25 ng/dL.

In one embodiment, this invention provides a method of lowering freeserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound of formula IA, I-XII orits isomer, pharmaceutical acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof, wherein the lowering offree serum testosterone levels is independent of a reduction of serumluteinizing hormone levels. In another embodiment the male subject hasprostate cancer.

In one embodiment, this invention provides methods of lowering totalserum testosterone or free serum testosterone levels in a male subject,wherein said male subject has prostate cancer. In another embodimentsaid subject has advanced prostate cancer.

In one embodiment, the reduction in serum concentrations of testosteroneis reversible and return to base line levels after treatment with thecompounds of this invention.

In another embodiment, serum concentrations of testosterone arereversible after treatment with Compound IV according to FIG. 23 andExample 10.

In one embodiment, this invention provides methods of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound of formula IA, I-XII orits isomer, pharmaceutical acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof. In another embodiment,the total serum testosterone is lowered below about 100 ng/dL. Inanother embodiment, the total serum testosterone is lowered below about50 ng/dL. In another embodiment, the total serum testosterone is loweredbelow about 25 ng/dL. In another embodiment, the total serumtestosterone is lowered below about 75 ng/dL. In another embodiment, thetotal serum testosterone is lowered to about between 75 ng/dL-100 ng/dL.In another embodiment, the total serum testosterone is lowered to aboutbetween 50 ng/dL-75 ng/dL. In another embodiment, the total serumtestosterone is lowered to about between 40 ng/dL-50 ng/dL. In anotherembodiment, the total serum testosterone concentration is lowered toabout between 25 ng/dL-50 ng/dL. In another embodiment, the total serumtestosterone is lowered to about between 40 ng/dL-60 ng/dL.

Testosterone can be measured as “free” (that is, bioavailable andunbound) or as “total” (including the percentage which is protein boundand unavailable) serum levels. In one embodiment, total serumtestosterone comprises free testosterone and bound testosterone.

Men, without prostate cancer, older than 40 years demonstrate lowtestosterone levels having total testosterone level of less than 250ng/dL (<8.7 nmol/L) or a free testosterone level of less than 0.75 ng/dL(<0.03 nmol/L). Methods of this invention provide a method of loweringserum testosterone levels. In one embodiment, methods provided lowertotal serum testosterone. In another embodiment, methods provided lowerfree serum testosterone.

In one embodiment, the methods of this invention provides a method oflowering total serum and/or free testosterone levels independent fromreduction of luteinizing hormone (LH) levels or by reduction of LHlevels in a male subject having prostate cancer. In another embodimentchanges in testosterone levels should be a reduction from the levelprior to treatment. In another embodiment, the total serum testosteronelevel is lowered below 100 ng/dL. In another embodiment, the total serumtestosterone is lowered below 50 ng/dL. In another embodiment, the totalserum testosterone is lowered below 25 ng/dL. In another embodiment, thefree testosterone level is lowered below 2 ng/dL. In another embodiment,the free testosterone level is lowered below 1 ng/dL. In anotherembodiment, the free testosterone level is lowered below 0.5 ng/dL. Inanother embodiment, the free testosterone level is lowered below 0.25ng/dL.

Methods of determining the free serum testosterone levels and totalserum testosterone levels include monitoring the testosterone levelsduring the course of the treatment period by a blood test. Totaltestosterone is a combination of circulating testosterone bound tocarrier proteins (albumin, SHBG, transcortin, transferrin) and thefree/unbound hormone. Total testosterone levels may be affected byseveral factors including the level of protein in the blood thattransports the hormone in the body, age, obesity and interferencesassociated with commonly used test methods.

Methods available to measure free testosterone (FT) can be complex(equilibrium dialysis and calculated free testosterone (CFT)) or simple(the commercial FT kit “Coat-A-Count”) using an analog tracer. Inanother embodiment the measurement of total testosterone and freetestosterone serum levels can be achieved by simultaneous measurement oftotal testosterone and SHBG (e.g., Irma-Count, DPC) and then acalculated free testosterone (CFT). In another embodiment themeasurement of total testosterone and free testosterone is according tothe knowledge of one skilled in the art.

In one embodiment, this invention provides a method of lowering totalserum testosterone levels or free serum testosterone levels in a malesubject comprising administering a therapeutically effective amount of acombination of one or more other forms of ADT and a compound of formulaIA, I-XII or its isomer, pharmaceutical acceptable salt, pharmaceuticalproduct, polymorph, hydrate or any combination thereof. In anotherembodiment, lowering of total or free serum testosterone occurs by areduction of serum luteinizing hormone (LH) level. In anotherembodiment, lowering total or free serum testosterone levels isindependent of a reduction of serum luteinizing hormone levels.

The methods of this invention comprise administering a combination ofother forms of ADT and a compound of this invention. In one embodiment,other forms of ADT include a LHRH agonist. In another embodiment theLHRH agonist includes Leuprolide acetate (Lupron®) (U.S. Pat. No.5,480,656; U.S. Pat. Nos. 5,575,987; 5,631,020; 5,643,607; 5,716,640;5,814,342; 6,036,976 which are all incorporated by reference herein) orgoserelin acetate (Zoladex®) (U.S. Pat. Nos. 7,118,552; 7,220,247;7,500,964 which are all incorporated by reference herein). In oneembodiment, other forms of ADT include an LHRH antagonist. In anotherembodiment, the LHRH antagonist includes degarelix. In one embodiment,other forms of ADT include anti-androgens. In another embodiment theanti-androgens include bicalutamide, flutamide, finasteride,dutasteride, MDV3100, nilutamide, chlormadinone, or any combinationthereof.

In one embodiment, the methods of this invention comprise administeringa therapeutically effective amount of an anti-androgen and a compound ofthis invention. In one embodiment, the methods of this inventioncomprise administering a therapeutically effective amount of an LHRHagonist and a compound of this invention. In one embodiment, the methodsof this invention comprise administering a therapeutically effectiveamount of an anti-androgen, LHRH agonist and a compound of thisinvention.

In one embodiment, this invention provides a method for lowering totalserum testosterone levels and/or free testosterone levels by reductionof luteinizing hormone (LH) levels or independent of reduction ofluteinizing hormone levels in a male subject having prostate cancer forthe purpose of producing androgen deprivation therapy (ADT) comprisingadministering a therapeutically effective amount of a compound offormula IA, I-XII. In another embodiment, the compound is Compound IV.

In another embodiment, this invention provides a method for androgendeprivation therapy (ADT) in a subject, comprising administering atherapeutically effective amount of a compound of formula IA, I-XII orits isomer, pharmaceutical acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof. In another embodiment,said subject has prostate cancer. In another embodiment, the compound isCompound IV.

In another embodiment, ADT is used for treating prostate cancer, fordelaying the progression of prostate cancer, or for preventing and/ortreating the recurrence of prostate cancer.

In one embodiment, this invention provides a method of treating prostatecancer or delaying the progression of prostate cancer comprisingadministering a compound of this invention. In one embodiment, thisinvention provides a method of preventing and/or treating the recurrenceof prostate cancer comprising administering a compound of thisinvention. In one embodiment, this invention provides a method ofincreasing the survival of a subject having prostate cancer, advancedprostate cancer, or castration-resistant prostate cancer comprisingadministering a compound of this invention. In another embodiment,administering a compound of this invention in combination with LHRHanalogs, reversible anti-androgens (such as bicalutamide or flutamide),anti-estrogens, anticancer drugs, 5-alpha reductase inhibitors,aromatase inhibitors, progestins, selective androgen receptor modulators(SARMs) or agents acting through other nuclear hormone receptors.

In one embodiment, the present invention provides a method of treatingprostate cancer and reducing of total serum testosterone and/or freeserum testosterone levels, by reducing LH levels or independent ofreduction of LH levels, comprising administering a compound of formulaIA, I-XII. In another embodiment, administering Compound IV.

Androgen deprivation therapy not only reduces testosterone, but estrogenlevels are also lower as estrogen is derived from the aromatization oftestosterone. Androgen deprivation therapy-induced estrogen deficiencycauses significant side effects which include hot flushes, gynecomastiaand mastalgia, bone loss, decreases in bone quality and strength,osteoporosis, osteopenia, and life-threatening fractures, adverse lipidchanges and higher cardiovascular disease and myocardial infarction,loss of libido, impotence, loss of muscle mass (sarcopenia), fatigue,cognitive dysfunction, and depression and other mood changes.

In other embodiments, the present invention provides a method oftreating any disease, disorder, or symptom associated with ADT. In otherembodiments, the present invention provides a method of treating anydisease, disorder, or symptom associated with testosterone deprivation.Each disease, disorder, or symptom represents a separate embodiment ofthe present invention.

In one embodiment, this invention provides a method of lowering totalserum testosterone levels in a male subject comprising administering atherapeutically effective amount of a compound of formulas IA, I-XII orits isomer, pharmaceutical acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof, wherein saidadministering said compounds of formulas IA, I-XII or its isomer,pharmaceutical acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof, prevents, suppresses, reduces theincidence, inhibits or treats side effects associated with androgendeprivation therapy (ADT) from occurring, wherein said subject hasprostate cancer. In another embodiment the lowering of the total or freeserum testosterone levels is by reducing LH levels or is independent ofreduction of LH levels.

In one embodiment, administering the compounds of this inventionsuppresses, reduces the incidence, inhibits or treats typical sideeffects associated with traditional androgen deprivation therapy (ADT)from occurring. In another embodiment, the subject has prostate cancer.Such prevention and/or reduction of side effects are relative to placeboor control group. In one embodiment, the typical side effects associatedwith traditional androgen deprivation therapy (ADT) include hot flashes,gynecomastia, decreased bone mineral density and increased bonefracture. In another embodiment, administering the compounds of thisinvention prevents hot flashes from occurring as would be found usingtraditional forms of androgen deprivation therapy (ADT). In anotherembodiment, administering the compounds of this invention preventsgynecomastia from occurring as would be found using traditional forms ofandrogen deprivation therapy (ADT). In another embodiment, administeringthe compounds of this invention prevents decreased bone mineral density(BMD) from occurring as would be found using traditional forms ofandrogen deprivation therapy (ADT). In another embodiment, administeringthe compounds of this invention prevents increased bone fracture fromoccurring as would be found using traditional forms of androgendeprivation therapy (ADT). In another embodiment, increased bonefracture is pathological fractures, non-traumatic fractures, vertebralfracture, non-vertebral fractures, new morphometric fractures, clinicalfracture or a combination thereof.

In one embodiment, administering the compounds of this invention lowerstotal serum testosterone without causing typical side effects associatedwith traditional androgen deprivation therapy (ADT) from occurring. Inanother embodiment, the subject has prostate cancer. In yet anotherembodiment, the subject has advanced prostate cancer. In still anotherembodiment, the subject has CRPC. In one embodiment, the typical sideeffects associated with traditional androgen deprivation therapy (ADT)include hot flashes, gynecomastia, decreased bone mineral density andincreased bone fracture. In another embodiment, the typical side effectassociated with traditional ADT includes increased body fat. In anotherembodiment, administering the compounds of this invention does not causehot flashes to occur as would be found using traditional forms ofandrogen deprivation therapy (ADT). In another embodiment, administeringthe compounds of this invention does not cause gynecomastia to occur aswould be found using traditional forms of androgen deprivation therapy(ADT). In another embodiment, administering the compounds of thisinvention does not cause decreased bone mineral density (BMD) to occuras would be found using traditional forms of androgen deprivationtherapy (ADT). In another embodiment, administering the compounds ofthis invention does not cause increased bone fracture to occur as wouldbe found using traditional forms of androgen deprivation therapy (ADT).In another embodiment, increased bone fracture is pathologicalfractures, non-traumatic fractures, vertebral fracture, non-vertebralfractures, new morphometric fractures, clinical fracture or acombination thereof. In yet another embodiment, administering thecompounds of this invention does not cause increased body fat to occuras would be found using traditional forms of androgen deprivationtherapy (ADT).

In one embodiment, administering the compounds of this invention lowersfree testosterone without causing typical side effects associated withtraditional androgen deprivation therapy (ADT) from occurring. Inanother embodiment, the subject has prostate cancer. In yet anotherembodiment, the subject has advanced prostate cancer. In still anotherembodiment, the subject has CRPC. In one embodiment, the typical sideeffects associated with traditional androgen deprivation therapy (ADT)include hot flashes, gynecomastia, decreased bone mineral density andincreased bone fracture. In another embodiment, the typical side effectassociated with traditional ADT includes increased body fat. In anotherembodiment, administering the compounds of this invention does not causehot flashes to occur as would be found using traditional forms ofandrogen deprivation therapy (ADT). In another embodiment, administeringthe compounds of this invention does not cause gynecomastia to occur aswould be found using traditional forms of androgen deprivation therapy(ADT). In another embodiment, administering the compounds of thisinvention does not cause decreased bone mineral density (BMD) to occuras would be found using traditional forms of androgen deprivationtherapy (ADT). In another embodiment, administering the compounds ofthis invention does not cause increased bone fracture to occur as wouldbe found using traditional forms of androgen deprivation therapy (ADT).In another embodiment, increased bone fracture is pathologicalfractures, non-traumatic fractures, vertebral fracture, non-vertebralfractures, new morphometric fractures, clinical fracture or acombination thereof. In yet another embodiment, administering thecompounds of this invention does not cause increased body fat to occuras would be found using traditional forms of androgen deprivationtherapy (ADT).

In one embodiment, the term “hot flashes” refers to sudden feeling ofheat in the upper part or all of the body, face and neck flush, redblotches appearing on the chest, back and arms, heavy sweating, coldshivering, etc.

In one embodiment, the term “gynecomastia” refers to a benignenlargement of the male breast resulting from a proliferation of theglandular component of the breast, which may or may not be associatedwith pain. Gynecomastia is defined clinically by the presence of arubbery or firm mass extending concentrically from the nipples. Thecondition known as pseudogynecomastia, or lipomastia, is characterizedby fat deposition without glandular proliferation. Although gynecomastiais usually bilateral, it can be unilateral.

In one embodiment, the methods of this invention are directed totreating men with prostate cancer or advanced prostate cancer orcastration-resistant prostate cancer by reduction of testosteronewithout also causing bone loss and hot flashes. In one embodiment, themethods of this invention are directed to treating men with prostatecancer or advanced prostate cancer or castration-resistant prostatecancer without also causing bone loss, gynecomastia and hot flashes.

In another embodiment, the methods of this invention make use ofcompounds IA, I-XII, wherein the compounds has the potential to reducetestosterone, a primary stimulus for prostate cancer, without alsocausing certain side effects such as bone loss and hot flashes which arecommon with current androgen deprivation therapies (ADT) for prostatecancer.

In another embodiment, Table 8 (Example 11) hereinbelow demonstratereduction of testosterone without also causing bone loss byadministering Compound IV.

In one embodiment, the methods of this invention are directed toreduction of testosterone levels which further treats advanced prostatecancer by administering a compound of formula IA, I-XII.

In one embodiment, the methods of this invention are directed toreduction of testosterone levels which further treatscastration-resistant prostate cancer by administering compound byadministering a compound of formulas IA, I-XII. In another embodiment,the methods of this invention are directed to reduction of testosteronelevels which further treats castration-resistant prostate cancer byadministering a compound of formula IA, I-XII.

In one embodiment, the methods of this invention are directed toreduction of testosterone levels which further suppresses, reduces theincidence, reduces the severity, or inhibits advanced prostate cancer byadministering a compound of formulas IA, I-XII. In another embodiment,the methods of this invention are directed to reduction of testosteronelevels which further suppresses, reduces the incidence, reduces theseverity, or inhibits advanced prostate cancer by administering compoundIV. In one embodiment, the methods of this invention are directed toreduction of testosterone levels which further provides palliativetreatment of advanced prostate cancer by administering a compound offormulas IA, I-XII. In another embodiment, the methods of this inventionare directed to reduction of testosterone levels which further providespalliative treatment of advanced prostate cancer by administeringcompound IV.

In one embodiment, the methods of this invention are directed totreating advanced prostate cancer. In another embodiment, the methods ofthis invention are directed to suppressing, reducing the incidence,reducing the severity, or inhibiting advanced prostate cancer. In oneembodiment, the methods of this invention are directed to palliativetreatment of advanced prostate cancer. In another embodiment, thisinvention is directed to suppressing advanced prostate cancer. Inanother embodiment, this invention is directed to reducing the incidenceof advanced prostate cancer. In another embodiment, this invention isdirected to reducing the severity of advanced prostate cancer. Inanother embodiment, this invention is directed to inhibiting advancedprostate cancer comprising administering a compound of this invention

In one embodiment, the methods of this invention are directed totreating castration-resistant prostate cancer. In one embodiment, themethods of this invention are directed to suppressing, reducing theincidence, reducing the severity, or inhibiting castration-resistantprostate cancer. In one embodiment, the methods of this invention aredirected to palliative treatment of castration-resistant prostatecancer. In another embodiment, this invention is directed to suppressingcastration-resistant prostate cancer. In another embodiment, thisinvention is directed to reducing the incidence of castration-resistantprostate cancer. In another embodiment, this invention is directed toreducing the severity of castration-resistant prostate cancer. Inanother embodiment, this invention is directed to inhibitingcastration-resistant prostate cancer. In another embodiment, thisinvention is directed to increase the survival of a subject withcastration-resistant prostate cancer. In another embodiment, the methodsof this invention make use of a compound of formulas IA, I-XII. Inanother embodiment, the methods of this invention make use of a compoundIV. In another embodiment, the methods of this invention make use of acompound of formulas IA, I-XII in combination with LHRH agonist. Inanother embodiment, the methods of this invention make use of compoundIV in combination with LHRH agonist. In another embodiment, the methodsof this invention make use of compound IV in combination with Leuprolideacetate (Lupron®). In another embodiment, the methods of this inventionmake use of a compound of formulas IA, I-XII in combination withLeuprolide acetate (Lupron®). In another embodiment, the methods of thisinvention make use of a compound of formulas IA, I-XII in combinationwith an anti-androgen. In another embodiment, the methods of thisinvention make use of compound IV in combination with an anti-androgen.

In another embodiment, this invention is directed to increase thesurvival of a subject with advanced prostate cancer. In anotherembodiment, the methods of this invention make use of a compound offormulas IA, I-XII. In another embodiment, the methods of this inventionmake use of a compound of formulas IA, I-XII in combination with LHRHagonist. In another embodiment, the methods of this invention make useof a compound of formulas IA, I-XII in combination with Leuprolideacetate (Lupron®). In another embodiment, the methods of this inventionmake use of a compound of formulas IA, I-XII in combination with ananti-androgen. In another embodiment, the methods of this invention makeuse of compound IV in combination with an anti-androgen.

In another embodiment, this invention is directed to increase thesurvival of a subject with advanced prostate cancer. In anotherembodiment, the methods of this invention make use of compound IV. Inanother embodiment, the methods of this invention make use of compoundIV in combination with LHRH agonist. In another embodiment, the methodsof this invention make use of compound IV in combination with Leuprolideacetate (Lupron®).

The term “advanced prostate cancer” refers to metastatic cancer havingoriginated in the prostate, and having widely metastasized to beyond theprostate such as the surrounding tissues to include the seminal vesiclesthe pelvic lymph nodes or bone, or to other parts of the body. Prostatecancer pathologies are graded with a Gleason grading from 1 to 5 inorder of increasing malignancy. In another embodiment, patients withsignificant risk of progressive disease and/or death from prostatecancer should be included in the definition and that any patient withcancer outside the prostate capsule with disease stages as low as IIBclearly has “advanced” disease.

Men with advanced prostate cancer often receive treatment to block theproduction of androgens, which are male sex hormones that may helpprostate tumors grow. However, prostate cancers that initially respondto anti-androgen therapy eventually develop the ability to grow withoutandrogens. Such cancers are often referred to as hormone refractory,androgen independent, or castration resistant.

In one embodiment, the advanced prostate cancer is castration-resistantprostate cancer.

In another embodiment, castration-resistant prostate cancer (CRPC) is anadvanced prostate cancer which developed despite ongoing ADT and/orsurgical castration. In another embodiment, ADT refers to treatmentconsisting Leuprolide acetate (Lupron®).

The term “castration-resistant prostate cancer” (CRPC) refers toprostate cancer which is considered hormone refractory, androgenindependent or castration resistant.

In one embodiment, castration-resistant prostate cancer is defined asprostate cancer that continues to progress or worsen or adversely affectthe health of the patient despite prior surgical castration, continuedtreatment with gonadotropin releasing hormone agonists (e.g.,leuprolide) or antagonists (degarelix), antiandrogens (e.g.,bicalutamide, flutamide, MDV3100, ketoconazole, aminoglutethamide),chemotherapeutic agents (e.g., docetaxel, paclitaxel, cabazitaxel,adriamycin, mitoxantrone, estramustine, cyclophosphamide), kinaseinhibitors (imatinib (Gleevec®) or gefitinib (Iressa®)) or otherprostate cancer therapies (e.g., vaccines (sipuleucel-T (Provenge®),GVAX, etc.), herbal (PC-SPES) and lyase inhibitor (abiraterone)) asevidenced by increasing or higher serum levels of prostate specificantigen (PSA), metastasis, bone metastasis, pain, lymph nodeinvolvement, increasing size or serum markers for tumor growth,worsening diagnostic markers of prognosis, or patient condition.

Many early prostate cancers require androgens for growth, but advancedprostate cancers are often androgen-independent. In men withcastration-resistant prostate cancer, the tumor cells may have theability to grow in the absence of androgens (hormones that promote thedevelopment and maintenance of male sex characteristics).

In one embodiment, the term “androgen deprivation therapy” (ADT) or“traditional androgen deprivation therapy” is directed to orchiectomy(surgical castration) wherein the surgeon removes the testicles. Inanother embodiment, the term “androgen deprivation therapy” or“traditional androgen deprivation therapy” is directed to administeringluteinizing hormone-releasing hormone (LHRH) analogs: These drugs lowerthe amount of testosterone made by the testicles. Examples of LHRHanalogs available in the United States include leuprolide (Lupron®,Viadur®, Eligard®), goserelin (Zoladex®), triptorelin (Trelstar®), andhistrelin (Vantas®). In another embodiment, the term “androgendeprivation therapy” or “traditional androgen deprivation therapy” isdirected to administering anti-androgens: Anti-androgens block thebody's ability to use any androgens. Even after orchiectomy or duringtreatment with LHRH analogs, a small amount of androgens is still madeby the adrenal glands. Examples of anti-androgens drugs include MDV3100,flutamide (Eulexin®), bicalutamide (Casodex®), and nilutamide(Nilandron®). In another embodiment, the term “androgen deprivationtherapy” or “traditional androgen deprivation therapy” is directed toadministering luteinizing hormone-releasing hormone (LHRH) antagonistssuch as abarelix (Plenaxis®) or degarelix (Firmagon®) (a new LHRHantagonist that was approved for use by the FDA in 2008 to treatadvanced prostate cancer). In another embodiment, the term “androgendeprivation therapy” or “traditional androgen deprivation therapy” isdirected to administering 5α-reductase inhibitors such as finasteride(Proscar®) and dutasteride (Avodart®): 5α-reductase inhibitors block thebody's ability to convert testosterone to the more active androgen,5α-dihydrotestosterone (DHT). In another embodiment, the term “androgendeprivation therapy” or “traditional androgen deprivation therapy” isdirected to administering inhibitors of testosterone biosynthesis suchas ketoconazole (Nizoral®). In another embodiment, the term “androgendeprivation therapy” or “traditional androgen deprivation therapy” isdirected to administering estrogens such as diethylstilbestrol or17β-estradiol.

In one embodiment, the methods of this invention are directed totreating, suppressing, reducing the incidence, reducing the severity,inhibiting, providing palliative care, or increasing the survival ofprostate cancer in a subject. In one embodiment, the methods of thisinvention are directed to methods of treating, suppressing, reducing theincidence, reducing the severity, inhibiting, providing palliative care,or increasing the survival of advanced prostate cancer in a subject. Inone embodiment, the methods of this invention are directed to treating,suppressing, reducing the incidence, reducing the severity, inhibiting,providing palliative care, or increasing the survival ofcastration-resistant prostate cancer. In another embodiment, the subjecthas high prostate specific antigen (PSA) levels.

In one embodiment, levels of prostate specific antigen (PSA) considerednormal are age dependent. In one embodiment, levels of prostate specificantigen (PSA) considered normal are dependent on the size of a malesubject's prostate. In one embodiment, PSA levels in the range between2.5-10 ng/mL are considered “borderline high”. In another embodiment,PSA levels above 10 ng/mL are considered “high”.

In one embodiment, the rate of change or “PSA velocity” is high. In oneembodiment, a rate of change or “PSA velocity” greater than 0.75/year isconsidered high.

In one embodiment, this invention is directed to treatment of a subjectwith high or increasing PSA levels comprising administering a compoundof this invention. In one embodiment, this invention is directed totreatment of a subject with high or increasing PSA levels despiteongoing ADT or a history of ADT, surgical castration or despitetreatment with anti-androgens and/or LHRH agonist.

In one embodiment, this invention is directed to a method of reducingthe prostate specific antigen (PSA) levels in a subject, comprisingadministering a compound of this invention. In one embodiment, thisinvention is directed to a method of reducing the prostate specificantigen (PSA) levels in a subject, comprising administering a compoundof formulas IA, I-XII. In another embodiment, by administering compoundIV. In one embodiment, this invention is directed to a method ofreducing the prostate specific antigen (PSA) levels in a subject,comprising administering a compound of formulas IA, I-XII in combinationwith LHRH agonist. In another embodiment, administering compound IV incombination with LHRH agonist. In one embodiment, this invention isdirected to a method of reducing the prostate specific antigen (PSA)levels in a subject, comprising administering a compound of formulas IA,I-XII in combination with leuprolide acetate (Lupron®). In anotherembodiment, administering compound IV in combination with leuprolideacetate (Lupron®).

In one embodiment, this invention provides methods of treatingcastration-resistant prostate cancer using the compounds of thisinvention, thereby requiring reduced chemotherapy.

In one embodiment, this invention provides a method of treating,suppressing, reducing the incidence, reducing the severity, increasingthe survival, or inhibiting a chemotherapy-resistant prostate cancer. Inanother embodiment, the chemotherapy comprises treatment with docetaxelor paclitaxel.

In one embodiment, this invention provides a method of treating,suppressing, reducing the incidence, reducing the severity, increasingthe survival, or inhibiting a GnRH agonist-resistant prostate cancer. Inanother embodiment, GnRH agonist is leuprolide.

In one embodiment, this invention provides a method of treating,suppressing, reducing the incidence, reducing the severity, increasingthe survival, or inhibiting a GnRH antagonist (GRHA)-resistant prostatecancer. In another embodiment, GRHA agonist is degarelix.

In one embodiment, this invention provides a method of treating,suppressing, reducing the incidence, reducing the severity, increasingthe survival, or inhibiting an antiandrogen-resistant prostate cancer.In another embodiment, the antiandrogen is bicalutamide or flutamide.

In one embodiment, this invention provides a method of treating,suppressing, reducing the incidence, reducing the severity, increasingthe survival, or inhibiting a vaccines-resistant prostate cancer.

In one embodiment, this invention provides a method of treating,suppressing, reducing the incidence, reducing the severity, increasingthe survival, or inhibiting an abiraterone-resistant prostate cancer.

In one embodiment, the methods provided herein and/or utilizing thecompounds provided herein, are effective in providing feedback on thehypothalamus-pituitary-testicular axis (HPT axis). Feedback refers tothe ability of a substance produced in one organ or tissue to regulatethe activity of another organ or tissue that affects its own activity.In one embodiment, feedback on the hypothalamus-pituitary-testicularaxis (HPT axis) results in reduction of LH levels. In one embodiment,feedback on the hypothalamus-pituitary-testicular axis (HPT axis)results in reduction of total serum testosterone levels. In oneembodiment, feedback on the hypothalamus-pituitary-testicular axis (HPTaxis) results in reduction of free serum testosterone levels. In oneembodiment, feedback on the hypothalamus-pituitary-testicular axis (HPTaxis) results in reduction of serum, tissue or tumor levels ofandrogens.

The hypothalamic-pituitary-testicular (HPT) axis refers to the endocrinephysiologic system that regulates hormone levels in the Hypothalmus, thePituitary gland and the Testes. LHRH (luteinizing hormone releasinghormone) is released by the hypothalamus and stimulates the pituitary tosynthesize and secrete LH and FSH (gonadotropins). LH and FSH then acton the testes to stimulate testosterone and sperm production.Testosterone then has a direct negative feedback effect on hypothalamicLHRH secretion and an indirect negative feedback effect on pituitary LHand FSH production. Estrogens, androgens and serum proteins (e.g.,inhibin) also have a negative effect on LHRH secretion and secretion ofLH and FSH.

The pituitary gland is one gland that controls the level of testosteronein the body. When the testosterone level is low, the pituitary glandreleases the luteinizing hormone (LH). This hormone induces the testesto make more testosterone. The level of testosterone increases duringpuberty. The level of testosterone is the highest around age 20 to 40,and then gradually becomes less in older men. Women have a much smalleramount of testosterone in their bodies compared to men. But testosteroneplays an important role throughout the body in both men and women. Itaffects the brain, bone and muscle mass, fat distribution, the vascularsystem, energy levels, genital tissues, and sexual function. Most of thetestosterone in the blood is bound to a protein called sex hormonebinding globulin (SHBG) or to another serum protein called albumin.Testosterone that is not bound (or “free”) can also be clinicallydetermined.

In another embodiment, lowering total serum testosterone or free serumtestosterone levels independent of a reduction of serum luteinizinghormone levels is due to increase of sex hormone-binding globulin(SHBG). In another embodiment, lowering free testosterone levelsindependent of a reduction of serum luteinizing hormone levels is due toincrease of sex hormone-binding globulin (SHBG). In another embodiment,lowering total serum or free serum testosterone levels independent of areduction of serum luteinizing hormone (LH) levels is due to inhibitionof testosterone production or secretion by Leydig cells in testes. Inanother embodiment, lowering total serum or free serum testosteronelevels independent of a reduction of serum luteinizing hormone (LH)levels is due to decrease of adrenal steroidogenesis.

In one embodiment, the compounds as described herein and/or compositionscomprising the same may be used for reduction of luteinizing hormone(LH) levels. In another embodiment, the compounds and/or compositions ofthis invention may be used to reduce endogenous sex hormones.

Hydroxysteroid dehydrogenase (HSD) family members are involved in theconversion of circulating steroids. 17β-HSD5 converts androstenedione totestosterone and estrone to estradiol. In addition, it is also involvedin prostaglandin synthesis. In one embodiment the compounds of thisinvention inhibit HSD specifically 17β-hydroxysteroid dehydrogenase 5(17β-HSD5) inhibition. Such inhibition may be useful in ADT, bypreventing the peripheral/extragonadal testosterone synthesis which mayescape the HPT axis control and cause incomplete reduction of total orfree serum testosterone or allow locally elevated intracellulartestosterone levels, either of which could be detrimental in ADT.

Androgen deprivation therapy (ADT) achieved by LHRH agonist therapy,i.e., administering luteinizing hormone releasing hormone agonists(LHRH) or analogues thereof, results in an initial stimulation ofgonadotropin release from the pituitary and testosterone production fromthe testes (termed “flare reaction”), followed by decrease ofgonadotropin release and decrease of both testosterone and estrogenlevels. The “flare reaction” caused by LHRH agonist therapy has anegative impact on treatment of prostate cancer, due to the increase ofandrogen/testosterone levels. In addition, LHRH therapy has beenassociated with increased risk of diabetes and cardiovascular disease(Smith (2008) Current Prostate Reports. 6:149-154).

In an effort to overcome the flare effects of LHRH therapy, antiandrogenmonotherapy (bicalutamide, flutamide, chlormadinone), combinedLHRH/antiandrogen therapy approaches, and LHRH antagonists (degarelix)have been suggested (Suzuki et al., (2008) Int. J. Clin. Oncol. 13:401-410; Sharifi, N. et al., (2005) JAMA. 294(2): 238-244). Antiandrogenmonotherapy does not reduce androgen levels in a subject. Bicalutamideantiandrogen monotherapy was shown to be less effective than ADT inprostate cancer patients with bone metastases. In addition, adverseeffects observed with bicalutamide therapy include breast tenderness andbreast enlargement (gynecomastia and mastodynia). (Suzuki et al., ibid)Additional risk with antiandrogen therapy includes increased livertransaminases. (Sharifi et al. ibid).

In one embodiment, the present invention provides a reduction of LHlevels and thereby a reduction of total serum testosterone and/or freeserum testosterone levels, without production of the “flare” effect, andwhile overcoming the adverse effects associated with estrogen deficitcaused by testosterone reduction using traditional ADT methods.Methods/uses of the subject compounds provide tissue-selective estrogenactivities that provide maintenance of bone tissue (agonist effect onbone tissue), decreased thrombic potential and/or hot flushes and/orlesser or neutral effects on breast tissue than estradiol ordiethylstilbestrol.

In one embodiment compound IV shows agonist but no antagonistic effects(Examples 6 and 7) so compound IV would not cause increase ingonadotropins and testosterone.

In one embodiment, compound IV shows agonist activity (Examples 8-11)demonstrating a robust pharmacologic response for the reduction of serumhormones, testosterone and total androgens.

In one embodiment, the methods provided herein utilizing the compoundsand/or compositions provided herein, are effective in reducing oreliminating bone resorptive effects caused by reduction of LH usingtraditional forms of ADT. In one embodiment, the methods provided hereinand/or utilizing the compositions provided herein, are effective inreducing or eliminating bone resorptive effects caused by reduction oftestosterone levels using traditional forms of ADT. In one embodiment,the methods provided herein utilizing the compositions provided herein,are effective in reducing or eliminating bone resorptive effects causedby reduction of estrogen as a result of LH level reduction. In oneembodiment, the methods provided herein utilizing the compounds and/orcompositions provided herein, prevent bone resorptive effects associatedwith LH level reduction using traditional forms of ADT. In oneembodiment, the methods provided herein utilizing the compounds and/orcompositions provided herein, prevent bone loss associated withendogenous LH, testosterone and/or estradiol reduction using traditionalforms of ADT. In one embodiment, the methods provided herein utilizingthe compounds and/or compositions provided herein, increase bone massdensity (BMD) while providing LH level reduction. In one embodiment, themethods provided herein utilizing the compounds and/or compositionsprovided herein, increase percent bone volume while providing endogenousLH, testosterone and/or estradiol level reduction.

In some embodiments, this invention provides a method of avoiding and/orreducing thromboembolism by administering a compound of this inventionor its isomer, pharmaceutical product, polymorph, hydrate or anycombination thereof.

In one embodiment, the methods provided herein utilizing the compoundsand/or compositions provided herein, are effective in breast tissue. Inone embodiment, the methods provided herein utilizing the compoundsand/or compositions provided herein, provide LH level reduction whilepreventing gynecomastia associated with LH level reduction achieved bytraditional ADT.

In one embodiment, Example 13 discloses special toxicity studies whereinin vitro studies with human platelets showed that Compound IV had muchlower procoagulatory activity than DES. Thus, Compound IV, anER-selective agonist, should deliver the prostate cancer benefits of DESwith lesser risk of thrombotic events than DES, and also deliver thebenefits of an LHRH agonist or antagonist without causing bone loss, hotflash or adverse lipid profiles.

Diethylstilbestrol (DES) therapy alone or combined with other ADT showedDES prevented bone resorption in patients with prostate cancer. Althoughuse of DES has been promoted as a therapy for prostate cancer, effectsof DES on angiogenesis and malignancy are thought to be mediated by DESmetabolites and are not thought to act through the estrogen receptor. Inaddition, dosage levels of DES administered for therapeutic uses presentnumerous adverse side effects including vascular disease, cardiovascularmorbidity, thrombotic toxicity, gynecomastia, erectile dysfunction anddecreased libido (Schen and Pitts, ibid and Presti, J. C. Jr. (1996)JAMA. 275(15): 1153-6).

In one embodiment, the present invention overcomes the negative sideeffects of LHRH agonist or antagonist therapy, alone or in combinationwith anti-androgens or DES. In another embodiment, methods of thesubject invention provide androgen deprivation therapy without adverseestrogen deprivation side-effects, such as adverse bone relatedconditions, and without adverse estrogen stimulation side-effects, suchas gynecomastia. In another embodiment, methods of the current inventionprovide for a reduction of LH levels and thereby a reduction of totaland/or free serum testosterone levels, without production of the “flare”effect, while overcoming the adverse effects associated with estrogendeficit caused by LH reduction and overcoming the adverse effectsassociated with a general estrogen agonist increase observed with DEStherapy. Methods/uses of the subject compounds provide tissue-selectiveestrogen activities thereby providing maintenance of bone tissue(agonist effect on bone tissue), decreased thrombic potential andneutral effects on breast tissue.

Antiestrogenic effects of traditional selective estrogen receptormodulators (SERMs) such as tamoxifen, toremifene and raloxifene at thehypothalamic level result in an increase of gonadotropin levels or anincrease of LH levels in men, and thereby potentially resulting in anincrease in the testosterone serum levels. (Tsouri et al., 2008,Fertility and Sterility doi: 10.1016) In contrast, the methods of thisinvention provide reduction of LH in a male subject comprisingadministering a compound of formula IA, I-XII.

Additional Embodiments for Compound of Formula I

In one embodiment of the methods of this invention, Y of compound offormula I is C(O). In another embodiment Y is CH₂. In another embodimentR₁ and R₂ of the compound of formula I or IA are independentlyO-Alk-NR₅R₆ or O-Alk-heterocycle. In another embodiment the Alk of saidO-Alk-heterocycle, O-Alk-NR₅R₆, -Alk-heterocycle and Alk-NR₅R₆ asdescribed herein above are linear alkyl of 1-7 carbons, branched alkylof 1-7 carbons, or cyclic alkyl of 3-8 carbons. In another embodiment,the alkyl is ethylene (—CH₂CH₂—). In another embodiment the Alk ismethylene (—CH₂—). In another embodiment the Alk is propylene(—CH₂CH₂CH₂—). In another embodiment the Alk is 2-methylpropylene(—CH₂CH(CH₃)CH₂—).

In one embodiment of the methods of this invention R₁ of the compound offormula I or IA is in the para position. In one embodiment of themethods of this invention R₁ and R₂ of the compound of formula I or IAare different. In another embodiment of the methods of this invention R₁and R₂ of the compound of formula I or IA are the same. In anotherembodiment of the methods of this invention R₁ of the compound offormula I or IA is

In another embodiment of the methods, R₁ of the compound of formula I orIA is hydroxyl. In another embodiment of the methods, R₁ of the compoundof formula I or IA is alkoxy. In another embodiment of the methods, R₁and R₂ are independently hydrogen, halogen, hydroxyl, alkoxy, cyano,nitro, CF₃, N(R)₂, sulfonamide, SO₂R, alkyl, haloalkyl, aryl,O-Alk-NR₅R₆ or O-Alk-heterocycle in which the heterocycle is a 3-7membered substituted or unsubstituted heterocyclic ring, optionallyaromatic. In another embodiment of the methods, R₁ and R₂ of thecompound of formula I or IA are independently halogen, hydroxyl, alkoxy,cyano, nitro, CF₃, N(R)₂, sulfonamide, SO₂R, alkyl, haloalkyl, aryl,O-Alk-NR₅R₆ or O-Alk-heterocycle in which the heterocycle is a 3-7membered substituted or unsubstituted heterocyclic ring, optionallyaromatic. In another embodiment of the methods, R₂ of the compound offormula I or IA is halogen. In another embodiment of the methods, R₂ ofthe compound of formula I or IA is F. In another embodiment of themethods, R₂ of the compound of formula I is Cl. In another embodiment ofthe methods, R₂ of the compound of formula I or IA is Br. In anotherembodiment of the methods, R₂ of the compound of formula I or IA is I.In another embodiment of the methods, R₂ of the compound of formula I orIA is hydroxyl. In another embodiment of the methods, R₁ and/or R₂ isCF₃. In another embodiment, R₁ and/or R₂ is CH₃. In another embodiment,R₁ and/or R₂ is halogen. In another embodiment, R₁ and/or R₂ is F. Inanother embodiment, R₁ and/or R₂ is Cl. In another embodiment, R₁ and/orR₂ is Br. In another embodiment, R₁ and/or R₂ is I. In anotherembodiment, R₂ of compound of formula I is in the para position.

In one embodiment of the methods of this invention, R₃ and R₄ of thecompound of formula I or IA are the same. In another embodiment of themethods of this invention, R₃ and R₄ of the compound of formula I or IAare different. In another embodiment of the methods, j and k of thecompound of formula I or IA are independently 1. In another embodimentof the methods, R₃ and R₄ of the compound of formula I or IA areindependently halogen, haloalkyl, hydroxyl or alkyl. In anotherembodiment of the methods, R₃ and R₄ of the compound of formula I or IAare independently F. In another embodiment of the methods, R₃ and R₄ ofthe compound of formula I or IA are independently Br. In anotherembodiment of the methods, R₃ and R₄ of the compound of formula I or IAare independently Cl. In another embodiment, R₄ is in the para position.In another embodiment, R₃ is in the ortho position. In anotherembodiment, R₃ is in the meta position. In another embodiment, R₃ and/orR₄ is CF₃. In another embodiment, R₃ and/or R₄ is CH₃.

In one embodiment of the methods of this invention, R₅ and R₆ of thecompound of formula I or IA form a 3 to 7 membered ring with thenitrogen atom. In another embodiment the ring is saturated orunsaturated ring. In another embodiment the ring substituted orunsubstituted ring. In another embodiment of the methods of thisinvention, R₅ and R₆ of the compound of formula I or IA form apiperidine ring with the nitrogen. In another embodiment of the methods,R₅ and R₆ of the compound of formula I or IA form a pyrazine ring withthe nitrogen. In another embodiment of the methods, R₅ and R₆ of thecompound of formula I or IA form a piperazine ring with the nitrogen. Inanother embodiment of the methods, R₅ and R₆ of the compound of formulaI or IA form a morpholine ring with the nitrogen. In another embodimentof the methods, R₅ and R₆ of the compound of formula I or IA form apyrrole ring with the nitrogen. In another embodiment of the methods, R₅and R₆ of the compound of formula I or IA form a pyrrolidine. In anotherembodiment of the methods, R₅ and R₆ of the compound of formula I or IAform a pyridine ring with the nitrogen. In another embodiment the ringis substituted by halogen, alkyl, alkoxy, alkylene, hydroxyl, cyano,nitro, amino, amide, COOH or an aldehyde.

In another embodiment of the methods of this invention, R₁ of thecompound of formula I or IA and R₂ of compound of the compound offormula I or IA are independently O-Alk-heterocycle orOCH₂CH₂-heterocycle. In another embodiment, the term “heterocycle” grouprefers, in one embodiment, to a ring structure comprising in addition tocarbon atoms, sulfur, oxygen, nitrogen or any combination thereof, aspart of the ring. In another embodiment the heterocycle is a 3-12membered ring. In another embodiment the heterocycle is a 6 memberedring. In another embodiment the heterocycle is a 5-7 membered ring. Inanother embodiment the heterocycle is a 4-8 membered ring. In anotherembodiment, the heterocycle group may be unsubstituted or substituted bya halogen, haloalkyl, hydroxyl, alkoxy, carbonyl, amido, alkylamido,dialkylamido, cyano, nitro, CO₂H, amino, alkylamino, dialkylamino,carboxyl, thio and/or thioalkyl. In another embodiment, the heterocyclering may be fused to another saturated or unsaturated cycloalkyl orheterocyclic 3-8 membered ring. In another embodiment, the heterocyclicring is a saturated ring. In another embodiment, the heterocyclic ringis an unsaturated ring. In another embodiment, the heterocycle ispiperidine. In another embodiment, the heterocycle is pyridine. Inanother embodiment, the heterocycle is piperidine, pyridine, furan,thiophene, pyrrole, pyrrolidine, pyrazine, piperazine or pyrimidine.

The term “cycloalkyl” refers to a non-aromatic, monocyclic or polycyclicring comprising carbon and hydrogen atoms. A cycloalkyl group can haveone or more carbon-carbon double bonds in the ring so long as the ringis not rendered aromatic by their presence. Examples of cycloalkylgroups include, but are not limited to, (C3-C7) cycloalkyl groups, suchas cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl,and saturated cyclic and bicyclic terpenes and (C3-C7) cycloalkenylgroups, such as cyclopropenyl, cyclobutenyl, cyclopentenyl,cyclohexenyl, and cycloheptenyl, and unsaturated cyclic and bicyclicterpenes. A cycloalkyl group can be unsubstituted or substituted by oneor two substituents. Preferably, the cycloalkyl group is a monocyclicring or bicyclic ring.

The term “alkyl” refers, in one embodiment, to a saturated aliphatichydrocarbon, including straight-chain, branched-chain and cyclic alkylgroups. In one embodiment, the alkyl group has 1-12 carbons. In anotherembodiment, the alkyl group has 1-7 carbons. In another embodiment, thealkyl group has 1-6 carbons. In another embodiment, the alkyl group has1-4 carbons. In another embodiment, the cyclic alkyl group has 3-8carbons. In another embodiment, the cyclic alkyl group has 3-12 carbons.In another embodiment, the branched alkyl is an alkyl substituted byalkyl side chains of 1 to 5 carbons. In another embodiment, the branchedalkyl is an alkyl substituted by haloalkyl side chains of 1 to 5carbons. The alkyl group may be unsubstituted or substituted by ahalogen, haloalkyl, hydroxyl, alkoxy carbonyl, amido, alkylamido,dialkylamido, nitro, amino, alkylamino, dialkylamino, carboxyl, thioand/or thioalkyl.

An “alkenyl” group refers, in another embodiment, to an unsaturatedhydrocarbon, including straight chain, branched chain and cyclic groupshaving one or more double bonds. The alkenyl group may have one doublebond, two double bonds, three double bonds, etc. In another embodiment,the alkenyl group has 2-12 carbons. In another embodiment, the alkenylgroup has 2-6 carbons. In another embodiment, the alkenyl group has 2-4carbons. Examples of alkenyl groups are ethenyl, propenyl, butenyl,cyclohexenyl, etc. The alkenyl group may be unsubstituted or substitutedby a halogen, hydroxy, alkoxy carbonyl, amido, alkylamido, dialkylamido,nitro, amino, alkylamino, dialkylamino, carboxyl, thio and/or thioalkyl.

An “aryl” group refers to an aromatic group having at least onecarbocyclic aromatic group or heterocyclic aromatic group, which may beunsubstituted or substituted by one or more groups selected fromhalogen, haloalkyl, hydroxy, alkoxy carbonyl, amido, alkylamido,dialkylamido, nitro, amino, alkylamino, dialkylamino, carboxy or thio orthioalkyl. Nonlimiting examples of aryl rings are phenyl, naphthyl,pyranyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyrazolyl, pyridinyl,furanyl, thiophenyl, thiazolyl, imidazolyl, isoxazolyl, and the like. Inone embodiment, the aryl group is a 4-8 membered ring. In anotherembodiment, the aryl group is a 4-12 membered ring(s). In anotherembodiment, the aryl group is a 6 membered ring. In another embodiment,the aryl group is a 5 membered ring. In another embodiment, the arylgroup is 2-4 fused ring system.

A “aldehyde” group refers, in one embodiment to an alkyl, or alkenylsubstituted by a formyl group, wherein the alkyl or alkenyl are asdefined hereinabove. In another embodiment, the aldehyde group is anaryl, or phenyl group substituted by a formyl group, wherein the aryl isas defined hereinabove. Examples of aldehydes are: formyl, acetal,propanal, butanal, pentanal, benzaldehyde. In another embodiment, thealdehyde group is a formyl group.

A “haloalkyl” group refers, in another embodiment, to an alkyl group asdefined above, which is substituted by one or more halogen atoms, e.g.by F, Cl, Br or I.

A “hydroxyl” group refers, in another embodiment, to an OH group. It isunderstood by a person skilled in the art that when R₁, R₂ or R₃ in thecompounds of the present invention is OR, then R is not OH.

In one embodiment, the term “halogen” or “halo” refers to a halogen,such as F, Cl, Br or I.

In another embodiment, the phrase “phenol” refers to an alcohol (OH)derivative of benzene.

Reference to protected hydroxyl, in some embodiments, includes theincorporation of a substituent bonded to the oxygen moiety of thebenzene ring, wherein the substituent may be readily removed. In someembodiments, phenolic protecting groups may comprise a: methyl ether(methoxy), alkyl ether (alkoxy), benzyl ether (Bn), methoxymethyl (MOM)ether, benzoyloxymethyl (BOM) ether, benzyl, carbobenzoxy,methoxyethoxymethyl (MEM) ether, 2-(trimethylsilyl)ethoxymethyl (SEM)ether, methylthiomethyl (MTM) ether, phenylthiomethyl (PTM) ether,azidomethyl ether, cyanomethyl ether, 2,2-dichloro-1,1-difluoroethylether, 2-chloroethyl ether, 2-bromoethyl ether, tetrahydropyranyl (THP)ether, 1-ethoxyethyl (EE) ether, phenacyl ether, 4-bromophenacyl ether,cyclopropylmethyl ether, allyl ether, propargyl ether, isopropyl ether,cyclohexyl ether, t-butyl ether, 2,6-dimethylbenzyl ether,4-methoxybenzyl ether, o-nitrobenzyl ether, 2,6-dichlorobenzyl ether,3,4-dichlorobenzyl ether, 4-(dimethylamino)carbonylbenzyl ether,4-methylsulfinylbenzyl ether, 4-anthrylmethyl ether, 4-picolyl ether,heptafluoro-p-tolyl, tetrafluoro-4-pyridyl ether, trimethylsilyl (TMS)ether, t-butyldimethylsilyl (TBDMS) ether, t-butyldiphenylsilyl (TBDPS)ether, triisopropylsilyl (TIPS) ether, aryl formate, arylacetate, aryllevulinate, arylpivaloate, aryl benzoate, aryl 9-fluorencarboxylate,aryl methyl carbonate, 1-adamantyl carbonate, t-butyl carbonate,4-methylsulfinylbenzyl carbonate, 2,4-dimethylpent-3-yl carbonate, aryl2,2,2-trichloroethyl carbonate, aryl benzyl carbonate, aryl carbamate,dimethylphosphinyl ester (Dmp-OAr), dimethylphosphinothionyl ester(Mpt-OAr), diphenylphosphinothionyl ester (Dpt-OAr), arylmethanesulfonate, aryl toluenesulfonate or aryl2-formylbenzenesulfonate.

In one embodiment, the methods of this invention make use ofN,N-bis(4-hydroxyphenyl)-4-propylbenzamide (II) or its isomer,pharmaceutically acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof. In another embodiment the methods ofthis invention make use of 4,4′-(2,3-dimethyl-benzylazanediyl)diphenol(III) or its isomer, pharmaceutically acceptable salt, pharmaceuticalproduct, polymorph, hydrate or any combination thereof. In anotherembodiment the methods of this invention make use of3-fluoro-N-(4-fluorophenyl)-4-hydroxy-N-(4-hydroxyphenyl)benzamide (IV)or its isomer, pharmaceutically acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof. In another embodiment themethods of this invention make use ofN,N-bis(4-hydroxyphenyl)-2,3-dimethylbenzamide (V) or its isomer,pharmaceutically acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof. In another embodiment the methods ofthis invention make use of N,N-bis(4-hydroxyphenyl)-2-naphthylamide (VI)or its isomer, pharmaceutically acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof. In another embodiment themethods of this invention make use of3-fluoro-4-hydroxy-N,N-bis(4-hydroxyphenyl)-benzamide (VII) or itsisomer, pharmaceutically acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof. In another embodiment themethods of this invention make use of a4-((4-fluorophenyl)(4-hydroxybenzyl)amino)phenol (VIII) or its isomer,pharmaceutically acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof. In another embodiment the methods ofthis invention make use of a4-fluoro-N-(4-hydroxy-phenyl)-N-[4-(2-piperidin-1-yl-ethoxy)-phenyl]-2-trifluoromethyl-benzamide(IX) or its isomer, pharmaceutically acceptable salt, pharmaceuticalproduct, polymorph, hydrate or any combination thereof. In anotherembodiment the methods of this invention make use of a hydrochloridesalt of IX (HCl salt of IX) or4-fluoro-N-(4-hydroxy-phenyl)-N-[4-(2-piperidin-1-yl-ethoxy)-phenyl]-2-trifluoromethyl-benzamidehydrochloride (X) or its isomer, pharmaceutically acceptable salt,pharmaceutical product, polymorph, hydrate or any combination thereof.In another embodiment the methods of this invention make use of a3-fluoro-4-hydroxy-N-(4-hydroxyphenyl)-N-phenylbenzamide (XI) or itsisomer, pharmaceutically acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof. In another embodiment themethods of this invention make use of a3-fluoro-N,N-bis-(4-hydroxy-phenyl)-2-methyl-benzamide (XII) or itsisomer, pharmaceutically acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof.

In one embodiment the methods of this invention make use of“pharmaceutically acceptable salts” of the compounds, which may beproduced, by reaction of a compound of this invention with an acid orbase.

Suitable pharmaceutically-acceptable salts of amines of the compounds ofthe methods of this invention may be prepared from an inorganic acid orfrom an organic acid. In one embodiment, examples of inorganic salts ofamines are bisulfates, borates, bromides, chlorides, hemisulfates,hydrobromates, hydrochlorates, 2-hydroxyethylsulfonates(hydroxyethanesulfonates), iodates, iodides, isothionates, nitrate,persulfates, phosphate, sulfates, sulfamates, sulfanilates, sulfonicacids (alkylsulfonates, arylsulfonates, halogen substitutedalkylsulfonates, halogen substituted arylsulfonates), sulfonates andthiocyanates.

In one embodiment, examples of organic salts of amines may be selectedfrom aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic,carboxylic and sulfonic classes of organic acids, examples of which areacetates, arginines, aspartates, ascorbates, adipates, anthranilates,algenates, alkane carboxylates, substituted alkane carboxylates,alginates, benzenesulfonates, benzoates, bisulfates, butyrates,bicarbonates, bitartrates, carboxylates, citrates, camphorates,camphorsulfonates, cyclohexylsulfamates, cyclopentanepropionates,calcium edetates, camsylates, carbonates, clavulanates, cinnamates,dicarboxylates, digluconates, dodecylsulfonates, dihydrochlorides,decanoates, enanthuates, ethanesulfonates, edetates, edisylates,estolates, esylates, fumarates, formates, fluorides, galacturonates,gluconates, glutamates, glycolates, glucorates, glucoheptanoates,glycerophosphates, gluceptates, glycollylarsanilates, glutarates,glutamate, heptanoates, hexanoates, hydroxymaleates, hydroxycarboxlicacids, hexylresorcinates, hydroxybenzoates, hydroxynaphthoates,hydrofluorates, lactates, lactobionates, laurates, malates, maleates,methylenebis(beta-oxynaphthoate), malonates, mandelates, mesylates,methane sulfonates, methylbromides, methylnitrates, methylsulfonates,monopotassium maleates, mucates, monocarboxylates, nitrates,naphthalenesulfonates, 2-naphthalenesulfonates, nicotinates, napsylates,N-methylglucamines, oxalates, octanoates, oleates, pamoates,phenylacetates, picrates, phenylbenzoates, pivalates, propionates,phthalates, pectinates, phenylpropionates, palmitates, pantothenates,polygalacturates, pyruvates, quinates, salicylates, succinates,stearates, sulfanilates, subacetates, tartarates, theophyllineacetates,p-toluenesulfonates (tosylates), trifluoroacetates, terephthalates,tannates, teoclates, trihaloacetates, triethiodide, tricarboxylates,undecanoates and valerates.

In one embodiment, examples of inorganic salts of carboxylic acids orphenols may be selected from ammonium, alkali metals to include lithium,sodium, potassium, cesium; alkaline earth metals to include calcium,magnesium, aluminium; zinc, barium, cholines, quaternary ammoniums.

In another embodiment, examples of organic salts of carboxylic acids orphenols may be selected from arginine, organic amines to includealiphatic organic amines, alicyclic organic amines, aromatic organicamines, benzathines, t-butylamines, benethamines(N-benzylphenethylamine), dicyclohexylamines, dimethylamines,diethanolamines, ethanolamines, ethylenediamines, hydrabamines,imidazoles, lysines, methylamines, meglamines, N-methyl-D-glucamines,N,N′-dibenzylethylenediamines, nicotinamides, organic amines,ornithines, pyridines, picolies, piperazines, procain,tris(hydroxymethyl)methylamines, triethylamines, triethanolamines,trimethylamines, tromethamines and ureas.

In one embodiment, the salts may be formed by conventional means, suchas by reacting the free base or free acid form of the product with oneor more equivalents of the appropriate acid or base in a solvent ormedium in which the salt is insoluble or in a solvent such as water,which is removed in vacuo or by freeze drying or by exchanging the ionsof a existing salt for another ion or suitable ion-exchange resin.

In one embodiment, the methods of this invention make use of apharmaceutically acceptable salt of the compounds of this invention. Inone embodiment the methods of this invention make use of apharmaceutically acceptable salt of compounds of formulas IA, I-XII. Inone embodiment, the methods of this invention make use of a salt of anamine of the compounds of formulas IA, I-XII of this invention. In oneembodiment, the methods of this invention make use of a salt of a phenolof the compounds of formulas IA, I-XII of this invention.

In one embodiment the methods of this invention make use of a free base,free acid, non charged or non-complexed compounds of formulas IA, I-XIIand/or its isomer, pharmaceutical product, hydrate, polymorph, orcombinations thereof.

In some embodiments of this invention, the compounds of this inventioncomprise three phenyl groups which are held together by an amide bond.In one embodiment, the compounds of this invention are non-chargedstructures. In another embodiment, the compounds of this invention arefree base structures. In another embodiment, the compounds of thisinvention are free acid structures. In another embodiment, the compoundsof this invention are non-complexed structures. In another embodiment,the compounds of this invention are non-ionized structures. In anotherembodiment, the compounds of this invention are pharmaceuticallyacceptable salts. In another embodiment, some compounds of thisinvention include hydrochloride (HCl) salts.

In one embodiment, the methods of this invention make use of an isomerof a compound of formulas IA, I-XII. In one embodiment, the methods ofthis invention make use of a pharmaceutical product of a compound offormulas IA, I-XII. In one embodiment, the methods of this inventionmake use of a hydrate of a compound of formulas IA, I-XII. In oneembodiment the methods of this invention make use of a polymorph of acompound of formulas IA, I-XII. In one embodiment the methods of thisinvention make use of a metabolite of a compound of formulas IA, I-XII.In another embodiment the methods of this invention make use of acomposition comprising a compound of formulas IA, I-XII, as describedherein, or, in another embodiment, a combination of isomer, metabolite,pharmaceutical product, hydrate, polymorph of a compound of formulas IA,I-XII.

In one embodiment, the term “isomer” includes, but is not limited to,optical isomers and analogs, structural isomers and analogs,conformational isomers and analogs, and the like.

In one embodiment, the term “isomer” is meant to encompass opticalisomers of the compound. In one embodiment, the term “isomer” is meantto encompass stereoisomers of the compound. The compounds of thisinvention possess an amide bond which may be in its cis or transisomerization. It is to be understood that the present inventionencompasses any optically-active, or stereroisomeric form, or mixturesthereof, and use of these for any application is to be considered withinthe scope of this invention.

In another embodiment, this invention further includes hydrates of thecompounds. In one embodiment, the term “hydrate” refers to hemihydrate,monohydrate, dihydrate, trihydrate or others, as known in the art.

Synthetic Processes

Compounds of Formula I or IA may readily be prepared, for example, byreacting a substituted diphenyl amine with benzoic acid or benzoylhalide in the presence of a base to yield a benzamide. In oneembodiment, the base is pyridine. In another embodiment, the benzoylhalide is benzoyl chloride. In another embodiment, a hydroxylsubstituent is protected during the reaction between the diphenylamineand the benzoic acid or benzoyl halide. In another embodiment, theprotecting group for the hydroxyl, optionally is removed in the laststep. See also U.S. Publication No. 2009/00624231, which is incorporatedby reference in its entirety.

For example, a compound of formula IA:

wherein R₁, R₂, R₃ and R₄, j and k are as described above;may be prepared by a process that comprises reacting

together with

to yield

andthe diphenyl amine (3) is reacted with

in the presence of a base to yield

wherein if R₁, R₂, R₃ and R₄ are independently OH, O-Alk-R₅R₆ orO-Alk-heterocycle, then R₁′, R₂′, R₃′, R₄′ are protected hydroxyl group,wherein the protecting group is removed to obtain the free hydroxyl oroptionally followed by reacting with Cl-Alk-heterocycle or Cl-Alk-NR₅R₆to yield a compound of formula IA:

wherein, if R₁, R₂, R₃ and R₄ are independently different than OH,O-Alk-NR₅R₆ or O-Alk-heterocycle then R₁′, R₂′, R₃′ and R₄′ are R₁, R₂,R₃ and R₄, respectively.

As another example, a process for the preparation of compound of FormulaIA:

wherein R₁, R₂, R₃ and R₄ are as described above, comprises reacting

with

in the presence of a base to yield

wherein if R₁, R₂, R₃ and R₄ are independently OH, O-Alk-R₅R₆ orO-Alk-heterocycle, then R₁′, R₂′, R₃′, R₄′ are protected hydroxyl group,wherein the protecting group is removed to obtain the free hydroxyl oroptionally followed by reacting with Cl-Alk-heterocycle or Cl-Alk-NR₅R₆to yield a compound of formula IA:

wherein, if R₁, R₂, R₃ and R₄ are independently different than OH,O-Alk-NR₅R₆ or O-Alk-heterocycle then R₁′, R₂′, R₃′ and R₄′ are R₁, R₂,R₃ and R₄, respectively.

In one example, Compound II is prepared according to Example 1, and FIG.5.

In another example Compound III is prepared according to Example 1, andFIG. 5.

In a further example a compound of formula IV (Compound IV):

may be prepared by reacting

with

in the presence of a base to yield

followed by deprotection of the protecting groups to yield Compound IV:

wherein P and P′ are the same or different protecting groups. In oneexample, Compound IV is prepared according to Example 2, and FIG. 6.

In another example, Compound V is prepared according to Example 1, andFIG. 5.

In a further example, Compound VI is prepared according to Example 3,and FIG. 7.

In another example, Compound VII is prepared according to Example 1, andFIG. 5.

In another example, Compound VIII is prepared according to Example 4,and FIG. 5.

In another example, Compound IX is prepared according to Example 5 andFIG. 8.

In another example, Compound X hydrochloride is prepared according toExample 5 and FIG. 8.

In another example, Compound XI is prepared according to Example 1, andFIG. 5.

In another example, Compound XII is prepared according to Example 1, andFIG. 5.

Suitable hydroxyl protecting groups include, for example, a methyl ether(methoxy), benzyl ether (benzyloxy)methoxymethyl (MOM) ether,benzoyloxymethyl (BOM) ether, benzyl, carbobenzoxy, methoxyethoxymethyl(MEM) ether, 2-(trimethylsilyl)ethoxymethyl (SEM) ether,methylthiomethyl (MTM) ether, phenylthiomethyl (PTM) ether, azidomethylether, cyanomethyl ether, 2,2-dichloro-1,1-difluoroethyl ether,2-chloroethyl ether, 2-bromoethyl ether, tetrahydropyranyl (THP) ether,1-ethoxyethyl (EE) ether, phenacyl ether, 4-bromophenacyl ether,cyclopropylmethyl ether, allyl ether, propargyl ether, isopropyl ether,cyclohexyl ether, t-butyl ether, benzyl ether, 2,6-dimethylbenzyl ether,4-methoxybenzyl ether, o-nitrobenzyl ether, 2,6-dichlorobenzyl ether,3,4-dichlorobenzyl ether, 4-(dimethylamino)carbonylbenzyl ether,4-methylsulfinylbenzyl ether, 4-anthrylmethyl ether, 4-picolyl ether,heptafluoro-p-tolyl, tetrafluoro-4-pyridyl ether, trimethylsilyl (TMS)ether, t-butyldimethylsilyl (TBDMS) ether, t-butyldiphenylsilyl (TBDPS)ether, triisopropylsilyl (TIPS) ether, aryl formate, arylacetate, aryllevulinate, arylpivaloate, aryl benzoate, aryl 9-fluorencarboxylate,aryl methyl carbonate, 1-adamantyl carbonate, t-butyl carbonate,4-methylsulfinylbenzyl carbonate, 2,4-dimethylpent-3-yl carbonate, aryl2,2,2-trichloroethyl carbonate, aryl benzyl carbonate, aryl carbamate,dimethylphosphinyl ester (Dmp-OAr), dimethylphosphinothionyl ester(Mpt-OAr), diphenylphosphinothionyl ester (Dpt-OAr), arylmethanesulfonate, aryl toluenesulfonate or aryl2-formylbenzenesulfonate.

The methods of this invention comprise the use of compounds of formulaIA or I-XII, wherein the process for the preparation of the compounds ofthis invention comprise reaction of a diphenyl amine with a benzoylchloride in the presence of a base. Suitable bases include, for example,pyridine, triethylamine, K₂CO₃, Cs₂CO₃, Na₂CO₃, methylamine, imidazole,benzimidazole, histidine, tributylamine or any combination thereof. Inone embodiment, the base is pyridine.

The methods of this invention comprise the use of compounds of formulaIA or I-XII, wherein the process for the preparation of the compounds ofthis invention comprises deprotection of a protected hydroxyl. Inanother embodiment, the deprotection conditions depend on the protectinggroup. In some embodiment, the deprotection step comprises hydrogenationin the presence of Pd/C. In another embodiment, the deprotectioncomprises reaction with BBr₃. In another embodiment, the deprotectionstep comprises reaction with an acid.

In further examples, Compounds of formula IA or I-XII are preparedaccording to FIGS. 5-8 and Examples 1-5.

Pharmaceutical Compositions

In some embodiments, this invention provides methods of use whichcomprise administering a composition comprising the described compounds.As used herein, “pharmaceutical composition” means a “therapeuticallyeffective amount” of the active ingredient, i.e. the compound of thisinvention, together with a pharmaceutically acceptable carrier ordiluent. A “therapeutically effective amount” as used herein refers tothat amount which provides a therapeutic effect for a given conditionand administration regimen.

As used herein, the term “administering” refers to bringing a subject incontact with a compound of the present invention. As used herein,administration can be accomplished in vitro, i.e. in a test tube, or invivo, i.e. in cells or tissues of living organisms, for example humans.In one embodiment, the present invention encompasses administering thecompounds of the present invention to a male subject.

This invention provides, in other embodiments, pharmaceutical productsof the compounds described herein. The term “pharmaceutical product”refers, in other embodiments, to a composition suitable forpharmaceutical use (pharmaceutical composition), for example, asdescribed herein.

The compounds of the invention can be administered alone or as an activeingredient of a formulation. Thus, the present invention also includespharmaceutical compositions of compounds of Formula I, containing, forexample, one or more pharmaceutically acceptable carriers.

Numerous standard references are available that describe procedures forpreparing various formulations suitable for administering the compoundsaccording to the invention. Examples of potential formulations andpreparations are contained, for example, in the Handbook ofPharmaceutical Excipients, American Pharmaceutical Association (currentedition); Pharmaceutical Dosage Forms Tablets (Lieberman, Lachman andSchwartz, editors) current edition, published by Marcel Dekker, Inc., aswell as Remington's Pharmaceutical Sciences (Arthur Osol, editor),1553-1593 (current edition).

The mode of administration and dosage forms is closely related to thetherapeutic amounts of the compounds or compositions which are desirableand efficacious for the given treatment application.

Suitable dosage forms include but are not limited to oral, rectal,sub-lingual, mucosal, nasal, ophthalmic, subcutaneous, intramuscular,intravenous, transdermal, spinal, intrathecal, intra-articular,intra-arterial, sub-arachinoid, bronchial, lymphatic, and intra-uterileadministration, and other dosage forms for systemic delivery of activeingredients. Formulations suitable for oral administration arepreferred.

To prepare such pharmaceutical dosage forms, the active ingredient maybe mixed with a pharmaceutical carrier according to conventionalpharmaceutical compounding techniques. The carrier may take a widevariety of forms depending on the form of preparation desired foradministration.

In preparing the compositions in oral dosage form, any of the usualpharmaceutical media may be employed. Thus, for liquid oralpreparations, such as, for example, suspensions, elixirs and solutions,suitable carriers and additives include water, glycols, oils, alcohols,flavoring agents, preservatives, coloring agents and the like. For solidoral preparations such as, for example, powders, capsules and tablets,suitable carriers and additives include starches, sugars, diluents,granulating agents, lubricants, binders, disintegrating agents and thelike. Due to their ease in administration, tablets and capsulesrepresent the most advantageous oral dosage unit form. If desired,tablets may be sugar coated or enteric coated by standard techniques.

For parenteral formulations, the carrier will usually comprise sterilewater, though other ingredients, for example, ingredients that aidsolubility or for preservation, may be included. Injectable solutionsmay also be prepared in which case appropriate stabilizing agents may beemployed.

In some applications, it may be advantageous to utilize the active agentin a “vectorized” form, such as by encapsulation of the active agent ina liposome or other encapsulant medium, or by fixation of the activeagent, e.g., by covalent bonding, chelation, or associativecoordination, on a suitable biomolecule, such as those selected fromproteins, lipoproteins, glycoproteins, and polysaccharides.

Treatment methods of the present invention using formulations suitablefor oral administration may be presented as discrete units such ascapsules, cachets, tablets, or lozenges, each containing a predeterminedamount of the active ingredient as, for example, a powder or granules.Optionally, a suspension in an aqueous liquor or a non-aqueous liquidmay be employed, such as a syrup, an elixir, an emulsion, or a draught.

A tablet may be made by compression or molding, or wet granulation,optionally with one or more accessory ingredients. Compressed tabletsmay be prepared by compressing in a suitable machine, with the activecompound being in a free-flowing form such as a powder or granules whichoptionally is mixed with, for example, a binder, disintegrant,lubricant, inert diluent, surface active agent, or discharging agent.Molded tablets comprised of a mixture of the powdered active compoundwith a suitable carrier may be made by molding in a suitable machine.

A syrup may be made by adding the active compound to a concentratedaqueous solution of a sugar, for example sucrose, to which may also beadded any accessory ingredient(s). Such accessory ingredient(s) mayinclude flavorings, suitable preservative, agents to retardcrystallization of the sugar, and agents to increase the solubility ofany other ingredient, such as a polyhydroxy alcohol, for exampleglycerol or sorbitol.

Formulations suitable for parenteral administration may comprise asterile aqueous preparation of the active compound, which preferably isisotonic with the blood of the recipient (e.g., physiological salinesolution). Such formulations may include suspending agents andthickening agents and liposomes or other microparticulate systems whichare designed to target the compound to blood components or one or moreorgans. The formulations may be presented in unit-dose or multi-doseform.

Parenteral administration may comprise any suitable form of systemicdelivery. Administration may for example be intravenous, intra-arterial,intrathecal, intramuscular, subcutaneous, intramuscular, intra-abdominal(e.g., intraperitoneal), etc., and may be effected by infusion pumps(external or implantable) or any other suitable means appropriate to thedesired administration modality.

Nasal and other mucosal spray formulations (e.g. inhalable forms) cancomprise purified aqueous solutions of the active compounds withpreservative agents and isotonic agents. Such formulations arepreferably adjusted to a pH and isotonic state compatible with the nasalor other mucous membranes. Alternatively, they can be in the form offinely divided solid powders suspended in a gas carrier. Suchformulations may be delivered by any suitable means or method, e.g., bynebulizer, atomizer, metered dose inhaler, or the like.

Formulations for rectal administration may be presented as a suppositorywith a suitable carrier such as cocoa butter, hydrogenated fats, orhydrogenated fatty carboxylic acids.

Transdermal formulations may be prepared by incorporating the activeagent in a thixotropic or gelatinous carrier such as a cellulosicmedium, e.g., methyl cellulose or hydroxyethyl cellulose, with theresulting formulation then being packed in a transdermal device adaptedto be secured in dermal contact with the skin of a wearer.

In addition to the aforementioned ingredients, formulations of thisinvention may further include one or more accessory ingredient(s)selected from, for example, diluents, buffers, flavoring agents,binders, disintegrants, surface active agents, thickeners, lubricants,preservatives (including antioxidants), and the like.

The formulations of the present invention can have immediate release,sustained release, delayed-onset release or any other release profileknown to one skilled in the art.

In one embodiment, this invention provides methods of a) lowering totalserum testosterone levels; b) lowering free serum testosterone levels byreduction of luteinizing hormone (LH) or independent of reduction of LHhormone in a male subject having prostate cancer comprisingadministering an oral composition comprising a compound of formulas IA,I-XII. In additional embodiments, the methods of this invention make useof an oral composition comprising a compound of formula II, formula III,formula IV, formula V, formula VI, formula VII, formula VIII, formulaIX, formula X, formula XI or formula XII.

In one embodiment, this invention provides a method of treating prostatecancer by reducing LH levels or independent of reduction of LH levels ina male subject having prostate cancer comprising administering an oralcomposition comprising a compound of formulas IA, I-XII. In additionalembodiments, this invention provides methods of treating prostate cancerby reducing LH levels or independent of reduction of LH levels in a malesubject having prostate cancer comprising administering an oralcomposition comprising a compound of formula II, formula III, formulaIV, formula V, formula VI, formula VII, formula VIII, formula IX,formula X, formula XI or formula XII.

It is to be understood that this invention encompasses any embodiment ofa compound as described herein, which in some embodiments is referred toas “a compound of this invention”.

In one embodiment, the methods of this invention may compriseadministration of a compound of this invention at various dosages. Inone embodiment, a compound of this invention is administered at a dosageof 1-1500 mg per day. In additional embodiments, a compound of thisinvention is administered at a dose of 1-10 mg per day, 3-26 mg per day,3-60 mg per day, 3-16 mg per day, 3-30 mg per day, 10-26 mg per day,15-60 mg, 50-100 mg per day, 50-200 mg per day, 150-300 mg per day,20-50 mg per day, 5-50 mg per day, 200-500 mg per day, 150-500 mg perday, 200-1000 mg per day, 300-1500 mg per day or 100-1000 mg per day.

In one embodiment, the methods of this invention may compriseadministration of a compound of this invention at various dosages. Inone embodiment, a compound of this invention is administered at a dosageof 3 mg. In additional embodiments, a compound of this invention isadministered at a dosage of 10 mg, 30 mg, 50 mg, 100 mg, 200 mg, 300 mg,450 mg, 500 mg, 600 mg, 900 mg, 1000 mg, 1500 mg, or 2000 mg.

In one embodiment, the methods of this invention may compriseadministration of a compound of this invention at various dosages. Inone embodiment, a compound of this invention is administered at a dosageof 0.1 mg/kg/day. In additional embodiments, a compound of thisinvention is administered at a dosage between 0.2 to 30 mg/kg/day, or0.2 mg/kg/day, 0.3 mg/kg/day, 1 mg/kg/day, 3 mg/kg/day, 5 mg/kg/day, 10mg/kg/day, 20 mg/kg/day or 30 mg/kg/day.

In one embodiment of the methods of this invention are provided for useof a pharmaceutical composition comprising a compound of formulas IA,I-XII. In additional embodiments, the methods of this invention areprovided for use of a pharmaceutical composition comprising a compoundof formula II, formula III, formula IV, formula V, formula VI, formulaVII, formula VIII, formula IX, formula X, formula XI or formula XII.

In certain embodiment, the pharmaceutical composition is a solid dosageform. In another embodiment, the pharmaceutical composition is a tablet.In another embodiment, the pharmaceutical composition is a capsule. Inanother embodiment, the pharmaceutical composition is a solution. Inanother embodiment, the pharmaceutical composition is a transdermalpatch.

In one embodiment, use of a compound of this invention or a compositioncomprising the same, will have utility in inhibiting, suppressing,enhancing or stimulating a desired response in a subject, as will beunderstood by one skilled in the art. In another embodiment, thecompositions may further comprise additional active ingredients, whoseactivity is useful for the particular application for which the compoundof this invention is being administered.

For administration to mammals, and particularly humans, it is expectedthat the physician will determine the actual dosage and duration oftreatment, which will be most suitable for an individual and can varywith the age, weight, genetics and/or response of the particularindividual.

In some embodiments, any of the compositions of this invention willcomprise a compound of this invention, in any form or embodiment asdescribed herein. In some embodiments, any of the compositions of thisinvention will consist of a compound of this invention, in any form orembodiment as described herein. In some embodiments, of the compositionsof this invention will consist essentially of a compound of thisinvention, in any form or embodiment as described herein. In someembodiments, the term “comprise” refers to the inclusion of theindicated active agent, such as the compound of this invention, as wellas inclusion of other active agents, and pharmaceutically acceptablecarriers, excipients, emollients, stabilizers, etc., as are known in thepharmaceutical industry. In some embodiments, the term “consistingessentially of” refers to a composition, whose only active ingredient isthe indicated active ingredient, however, other compounds may beincluded which are for stabilizing, preserving, etc. the formulation,but are not involved directly in the therapeutic effect of the indicatedactive ingredient. In some embodiments, the term “consisting essentiallyof” may refer to components which facilitate the release of the activeingredient. In some embodiments, the term “consisting” refers to acomposition, which contains the active ingredient and a pharmaceuticallyacceptable carrier or excipient.

It is to be understood that any use of any of the compounds as hereindescribed may be used in the treatment of any disease, disorder orcondition as described herein, and represents an embodiment of thisinvention. In one embodiment, the compounds are a free base, free acid,non charged or non-complexed compound.

The following examples are presented in order to more fully illustratethe preferred embodiments of the invention. They should in no way,however, be construed as limiting the broad scope of the invention.

EXAMPLES Example 1 General Synthesis Procedures for Compounds ofFormulas II-XII and Synthetic Intermediates

The organic solvents, surfactants and antioxidants, etc., they may beused in the compositions described herein are typically readilyavailable from commercial sources. For example, PEG-300, polysorbate 80,Captex™ 200, Capmul™ MCM C8 may be purchased, for example, from DowChemical Company (Midland, Mich.), ICI Americas, Inc (Wilmington, Del.)or Abitec Corporation (Janesville, Wis.).

The estrogen receptor ligands described herein may be prepared in anumber of ways well known to those skilled in the art. For example, theestrogen receptor ligands described herein may be prepared by thesynthetic methods described in U.S. Patent Application Publication Nos.2009/0062341, the disclosures of each of which are hereby incorporatedby reference in their entireties.

General Synthesis of N,N-bis Aryl Benzamide Derivatives

General synthesis of diarylanilines (FIG. 5). A mixture of arylamine(1.5 equivalent), aryl iodide (1 equivalent), K₂CO₃ (2 equivalents), CuI(0.1 equivalent) and L-proline (0.2 equivalent) were mixed together anddissolved in anhydrous DMSO at room temperature. Then, the reactionmixture was stirred and heated to 90° C. for 28 hours. The mixture wascooled to room temperature and hydrolyzed with water. EtOAc was added topartition the solution. The EtOAc layer was separated, washed withbrine, and dried over anhydrous MgSO₄. The solvent was removed underreduced pressure. The solid residue was purified by flash columnchromatography (silica gel) using 5% EtOAc/hexanes as eluent to affordthe corresponding diarylaniline.

Bis-(4-methoxyphenyl)amine (1a): pale-yellow solid, 73% yield. M.p.98.6-99.0° C. ¹H NMR (CDCl₃, 300 MHz) δ 6.93-6.81 (m, 8H), 5.37 (s, br,1H), 3.78 (s, 6H). MS m/z 228.4 (M−H)⁺

N-(4-Methoxyphenyl)-phenylamine (1b): pale-yellow solid, 70% yield. M.p.106.3-106.5° C. ¹H NMR (CDCl₃, 300 MHz) δ 7.24-7.18 (m, 3H), 7.08-7.06(m, 2H), 6.92-6.84 (m, 4H), 5.61 (s, br, 1H), 3.79 (s, 3H). MS m/z 200.1(M+H)⁺.

N-(4-Fluorophenyl)-N-4-methoxyphenylamine (1c): pale-yellow solid, 54%yield. M.p. 60.6-61.0° C. ¹H NMR (CDCl₃, 300 MHz) δ 7.01-6.83 (m, 8H),3.78 (s, 3H). MS m/z 217 (M)⁺.

N-(4-Benzyloxyphenyl)-N-4-methoxyphenylamine (1d): pale-yellow solid,54% yield. M.p. 108.0-108.4° C. ¹H NMR (CDCl₃, 300 MHz) δ 7.34-7.08 (m,5H), 6.90-6.81 (s, 3H), 3.78 (s, 3H). MS m/z 306 (M+H)⁺.

General Synthesis of Benzamides. A mixture of arylaniline (1equivalent), benzoyl chlorides (1.3 equivalents), and pyridine (6equivalents) was mixed together and dissolved in anhydrous THF at roomtemperature. The mixture was stirred and refluxed for 24 hours. Thereaction solution was cooled to room temperature, and hydrolyzed byaddition of 2 N HCl solution. The solution was extracted with ethylacetate. The organic layer was washed with a saturated aqueous NaHCO₃solution to remove excess acid, dried over anhydrous MgSO₄, filtered andconcentrated under reduced pressure. The residue was purified by flashcolumn chromatography using EtOAc/hexanes (3/7 v/v) to afford thecorresponding benzamide compounds.

3-Fluoro-N-(4-fluorophenyl)-4-methoxy-N-(4-methoxyphenyl)benzamide (2a):yellow solid, M.p. 54-56° C., ¹H NMR (CDCl₃/TMS) δ 7.24-7.11 (m, 4H),7.05-6.97 (m, 4H), 6.85-6.78 (m, 3H), 3.86 (s, 3H), 3.79 (s, 3H). MS(ESI) m/z 370.1 [M+H]⁺

4-Fluoro-N,N-bis(4-methoxyphenyl)-2-(trifluoromethyl)benzamide (2b):colorless oil, 84.2% yield. ¹H NMR (CDCl₃, 300 MHz) δ 7.34-7.26 (m, 4H),7.09-7.01 (m, 3H), 6.91 (d, 2H, J=8.7 Hz), 6.87 (d, 2H, J=8.7 Hz), 3.80(s, 3H), 3.71 (s, 3H). MS m/z 442.1 (M+Na)⁺.

4-Methoxy-N-(4-methoxyphenyl)-N-(4-fluorophenyl)-benzamide (2c): whitesolid, 97% yield, M.p. 133.5.0-134.5° C. ¹H NMR (CDCl₃, 300 MHz) δ8.11-6.66 (m, 15H), 3.74 (s, 3H), 3.73 (s, 3H). MS m/z 384 (M+H)⁺.

N-(4-Methoxyphenyl)-N-(4-benzyloxyphenyl)-2-naphthylamide (2d): whitesolid, 58% yield. M.p. 174.9-175.5° C. ¹H NMR (CDCl₃, 300 MHz) δ 8.04(s, 1H), 7.77-7.74 (m, 2H), 7.64-7.61 (m, 1H), 7.51-7.43 (m, 4H),7.40-7.31 (m, 4H), 7.13-7.10 m, 4H), 6.88-6.78 (m, 4H), 4.99 (s, 2H),3.74 (s, 3H). MS m/z 460 (M+H)⁺.

4-Fluoro-N,N-bis(4-methoxyphenyl)-2-(trifluoromethyl)benzamide (2e):colorless oil, 84.2% yield. ¹H NMR (CDCl₃, 300 MHz) δ 7.34-7.26 (m, 4H),7.09-7.01 (m, 3H), 6.91 (d, 2H, J=8.7 Hz), 6.87 (d, 2H, J=8.7 Hz), 3.80(s, 3H), 3.71 (s, 3H). MS m/z 442.1 (M+Na)⁺.

General Procedure for Demethylation of Benzamide Derivatives Using BBr₃.A methoxybenzamide compound was dissolved in dry CH₂Cl₂. BBr₃ (1.0 MCH₂Cl₂ solution) was added dropwise at 0° C. The reaction solution wasslowly warmed to room temperature and allowed to stir overnight at roomtemperature. The mixture was cooled to 0° C. in an ice bath andhydrolyzed by adding water. EtOAc was added to partition the solution.The organic layer was separated; the aqueous layer was extracted withEtOAc. The organic layer was washed with brine and dried over anhydrousMgSO₄. The solvent was removed under reduced pressure. The residue waspurified by flash column chromatography using CH₃OH/CH₂Cl₂ (1/9 v/v) toafford the corresponding phenolic compounds.

4-Fluoro-N,N-bis(4-hydroxyphenyl)-2-(trifluoromethyl)benzamide (3a):white solid, 92.5% yield. ¹H NMR (DMSO-d₆, 300 MHz) δ 9.55 (s, 1H), 9.53(s, 1H), 7.69-7.58 (m, 2H), 7.46-7.39 (m, 1H), 7.18 (d, 2H, J=8.7 Hz),6.93 (d, 4H, J=8.7 Hz), 7.03 (d, 2H, J=8.4 Hz), 6.78 (d, 2H, J=8.7 Hz),6.57 (d, 2H, J=8.7 Hz). MS m/z 392.1 (M+H)⁺.

The following compounds where synthesized as described herein above andcharacterized and summarized in Table 1:N,N-bis(4-hydroxyphenyl)-4-propylbenzamide (II);3-fluoro-N-(4-fluorophenyl)-4-hydroxy-N-(4-hydroxyphenyl)benzamide (IV);N,N-bis(4-hydroxyphenyl)-2,3-dimethylbenzamide (V);3-fluoro-4-hydroxy-N,N-bis(4-hydroxyphenyl)-benzamide (VII);3-fluoro-4-hydroxy-N-(4-hydroxyphenyl)-N-phenylbenzamide (XI); and3-fluoro-N,N-bis(4-hydroxyphenyl)-2-methylbenzamide (XII).

General Procedures for Debenzylation of Benzyloxyphenyl-benzamides.Compound was dissolved in EtOH in a 250 mL hydrogenation bottle. Pd/Cpowder (5% mol) was added to the solution. The reaction vessel wasmounted to a hydrogenation apparatus under 20 psi pressure hydrogen gas.The reaction was monitored by TLC until the disappearance of startingmaterial. Then, the solvent was removed under reduced pressure. Theresidue was purified by flash column chromatography withhexanes/EtOAc=3/2 v/v to afford the desired product.

The following compounds where synthesized as described herein above andcharacterized and summarized in Table 1:N,N-bis(4-hydroxyphenyl)-2-naphthylamide (VI).

General Procedures for Reduction of Deprotected Benzamides. Benzamidecompounds were dissolved in 20 mL anhydrous THF at room temperature.H₃B(SMe₂) was added via a syringe at room temperature under argon. Thereaction solution was stirred and heated to reflux for 6 hours. Then,the reaction was quenched by adding 10 mL of MeOH at 0° C. The solventwas removed under reduced pressure. The residue was subjected to flashcolumn chromatography (silica-gel, CH₂Cl₂/MeOH=9/1 v/v) to afford thedesired product.

The following compounds where synthesized as described herein above andcharacterized and summarized in Table 1:4,4′-(2,3-dimethylbenzylazanediyl)diphenol (III);4-((4-fluorophenyl)(4-hydroxybenzyl)amino)phenol (VIII).

General synthesis of O-(2-piperidin-1-ylethoxy)-benzamides andanalogues. To a solution of hydroxyphenyl containing benzamide analogue(1 equivalent) in acetone, K₂CO₃ (3 equivalents) andN-chloroethyl-piperidine hydrochloride salt (1.2 equivalents) wereadded. The solution was heated to reflux for 6 hours. The solution wasevaporated to dryness. The residue was hydrolyzed by adding water, andthen extracted with ethyl acetate. The organic layers were separated anddried over anhydrous MgSO₄. The solvent was removed under reducedpressure. The residue was purified by flash chromatography withmethylene chloride/methanol=9/1 v/v to give the desired compound. Thehydrochloride salts were prepared by adding HCl in Et₂O to the methanolsolution of the compounds followed by evaporation of solvents.

The following compounds where synthesized as described herein above andcharacterized and summarized in Table 1:4-fluoro-N-(4-hydroxyphenyl)-N-(4-(2-(piperidin-1-yl)ethoxy)phenyl)-2-(trifluoromethyl)benzamide(IX); and4-fluoro-N-(4-hydroxyphenyl)-N-(4-(2-(piperidin-1-yl)ethoxy)phenyl)-2-(trifluoromethyl)benzamidehydrochloride (X) which is the HCl salt of IX.

TABLE 1 Physical Characterization of Compounds of Formulas II-XII. Cmpd# Structure PHYSICAL CHARACTERIZATION II.

¹H NMR (DMSO-d₆, 300 MHz) δ 9.46 (s, 2H, 2 X OH), 7.27-7.26 (m, 2H,ArH), 7.06-7.04 (m, 2H, ArH), 6.99-6.97 (m, 4H, ArH), 6.66-6.65 (m, 4H,ArH), 2.50 (s, 2H, CH₂, overlapped with DMSO peak), 1.53-1.52 (m, 2H,CH₂), 0.82 (t, J = 7.33 Hz, 3H, CH₃). m/z 346.0 (M − H)⁻ III.

Tan foam, 41% yield. M.p. 147-150° C. ¹H NMR (DMSO-d₆), 300 MHz) δ 8.92(s, 2H), 7.07 (d, J = 7.33 Hz, 1H), 7.00-6.94 (m, 2H), 6.76-6.72 (m,4H), 6.63-6.59 (m, 4H), 4.72 (s, 2H), 2.23 (s, 3H), 2.16 (s, 3H). m/z320.2 (M + H)⁺ IV.

Tan solid, 92% yield. M.p. 110-112° C. ¹H NMR (DMSO-d₆, 300 MHz) δ 10.14(bs, 1H), 9.71 (bs, 1H), 7.26-7.11 (m, 5H), 7.05-6.99 (m, 3H), 6.78 (t,J = 8.61 Hz, 2H), 6.68 (d, J = 8.68 Hz, 2H). m/z 364.1 (M + Na)⁺ V.

¹H NMR (DMSO-d₆, 300 MHz) δ 9.47 (bs, 2H, 2 X OH), 7.18 (d, J = 8.30 Hz,2H, ArH), 7.06 (d, J = 7.08 Hz, 1H, ArH), 7.00-6.92 (m, 4H, ArH), 6.78(d, J = 8.30 Hz, 2H, ArH), 6.51 (d, J = 8.06 Hz, 2H, ArH), 2.22 (s, 3H,CH₃), 2.15 (s, 3H, CH₃). m/z 334.3 (M + H)⁺ VI.

white solid, 70% yield. M.p. 264.3-265.2° C. (decomposed). ¹H NMR(DMSO-d₆, 500 MHz) δ 9.46 (s, 2H), 7.98 (s, 1H), 7.85-7.75 (m, 2H),7.75- 7.73 (m, 2H), 7.54-7.48 (m, 2H), 7.45-7.43 (m, 1H), 7.05 (s, 4H),6.66 (s, 4H). m/z 356 (M + H)⁺ VII.

¹H NMR (DMSO-d₆, 300 MHz) δ 10.25 (bs, 1H, OH), 9.48 (bs, 2H, 2 X OH),7.12-6.95 (m, 6H, ArH), 6.80-6.65 (m, 5H, ArH). m/z 338.0 (M − H)⁻ VIII.

yellow oil, 92% yield. ¹H NMR (DMSO-d₆, 500 MHz) δ 9.29 (s, 1H), 9.24(s, 1H), 7.09 (d, 2H, J = 8.3 Hz), 6.98 (d, 2H, J = 9.0 Hz), 6.94-6.91(m, 2H), 6.73 (d, 2H, J = 9.0 Hz), 6.68-6.64 (m, 4H), 4.70 (s, 2H). m/z307.8 (M − H)⁻ IX. and X. (HCl salt of IX.)

white solid, 57.7% yield. ¹H NMR (DMSO-d₆, 300 MHz) δ 9.57 (s, 1H),7.71-7.68 (m, 2H), 7.47-7.44 (m, 1H), 7.28 (d, 1H, J = 9.0 Hz), 7.18 (d,1H, J = 8.7 Hz), 7.13 (d, 1H, J = 8.7 Hz), 7.05 (d, 1H, J = 8.4 Hz),6.97 (d, 1H, J = 9.0 Hz), 6.80-6.76 (m, 2H), 6.57 (d, 1H, J = 87. Hz),4.06 (t, 1H, J = 6.0 Hz), 3.93 (t, 1H, J = 6.0 Hz), 2.66 (t, 1H, J = 5.7Hz), 2.55 (t, 1H, J = 5.4 Hz), 2.44 (s, 2H), 2.36 (s, 2H), 1.49-1.37 (m,6H). m/z 501.0 (M − H)⁻ XI.

¹H NMR (DMSO-d₆, 300 MHz) δ 9.95 (bs, 1H, OH), 9.47 (bs, 2H, 2 X OH),7.02-6.95 (m, 6H, ArH), 6.75-6.72 (m, 1H, ArH), 6.68-6.66 (m, 4H, ArH).m/z 324.0 (M + H)⁺ XII.

Pale-red solid. 72.0% yield. M.p. >240° C. ¹H NMR (DMSO-d₆, 300 MHz) δ9.50 (bs, 2H), 7.19- 6.79 (m, 7H), 6.61 (d, J = 8.93 Hz, 2H), 6.53 (d, J= 7.79 Hz, 2H), 2.23 (s, 3H). m/z 336.0 (M − H)⁻.

Example 2 Synthesis of the Compound of Formula IV (FIG. 6)

Step 1: Synthesis of 4-fluoro-N-(4-methoxyphenyl)aniline (1c)

A mixture of 4-fluoroaniline (78.63 g, 0.708 mol), 4-iodoanisole (138.00g, 0.590 mol), anhydrous K₂CO₃ (122.23 g, 0.884 mol), CuI (11.23 g,58.96 mmol) and L-proline (13.58 g, 0.118 mol) was mixed together in adry 1 L three-necked round-bottomed flask fitted with a stirring bar, areflux condenser and an argon inlet. Anhydrous DMSO (300 mL) was addedat room temperature. The reaction mixture was stirred and heated to 90°C. for 20 hours under argon. Then, the mixture was cooled to roomtemperature and hydrolyzed with water (300 mL). EtOAc (200 mL) was addedto partition the solution. The EtOAc layer was separated. The aqueouslayer was extracted with 100 mL of EtOAc. The EtOAc layers werecombined, washed with brine (2×100 mL) and dried over anhydrous MgSO₄(50 g). The solvent was removed under reduced pressure. The brown oilresidue was purified by flash column chromatography (silica gel,Hexanes/EtOAc=9/1 v/v) to afford 4-fluoro-N-(4-methoxyphenyl)aniline(1c) as a yellow solid product, 99.70 g, 77.8% yield. M.p. 46-48° C. MS(ESI) m/z 218.1 [M+H]⁺, ¹H NMR (DMSO-d₆, 300 MHz) δ 7.77 (bs, 1H),7.03-6.98 (m, 4H), 6.93-6.82 (m, 4H), 3.70 (s, 3H).

Step 2: Synthesis of3-fluoro-N-(4-fluorophenyl)-4-methoxy-N-(4-methoxyphenyl)benzamide (2a)

4-Fluoro-N-(4-methoxyphenyl)aniline (1c) (90.78 g, 0.418 mol) and3-fluoro-4-methoxybenzoyl chloride (94.55 g, 0.501 mol) were mixedtogether and dissolved in anhydrous THF (200 mL) in a dry 1 Lthree-necked round-bottomed flask fitted with a stirring bar, a refluxcondenser and an argon inlet. Anhydrous pyridine (132.22 g, 1.672 mol)was added via a syringe at room temperature under argon. The reactionmixture was stirred and heated to reflux overnight. Then, the reactionmixture was cooled to room temperature and filtered to remove pyridinesalt. The solution was concentrated to remove THF solvent. The residueoil was washed with 200 mL of 2N HCl solution and extracted with ethylacetate (2×200 mL). The combined organic layer was washed with asaturated aqueous Na₂CO₃ solution (150 mL) to remove excess benzoylchloride and acid, dried over MgSO₄ (50 g), filtered, and concentratedunder reduced pressure to give an oil. The residue was purified by flashcolumn chromatography using silica-gel with CH₂Cl₂/acetone (50/1 v/v) toafford the pure corresponding benzamide compound as a yellow solid. M.p.54-56° C. MS (ESI) m/z 370.1 [M+H]⁺, ¹H NMR (CDCl₃/TMS) δ 7.24-7.11 (m,4H), 7.05-6.97 (m, 4H), 6.85-6.78 (m, 3H), 3.86 (s, 3H), 3.79 (s, 3H).

Step 3: Synthesis of3-fluoro-N-(4-fluorophenyl)-4-hydroxy-N-(4-hydroxyphenyl)benzamide (IV)

Compound3-fluoro-N-(4-fluorophenyl)-4-methoxy-N-(4-methoxyphenyl)benzamide (2a)(138.0 g, 0.374 mol) was dissolved in dry CH₂Cl₂ (600 mL) at roomtemperature under an argon. BBr₃ (374.75 g, 1.496 mol) was addeddropwise with stirring via a syringe at 0° C. in an ice-bath under anargon. The reaction solution was allowed to stir at room temperatureovernight. Then, the solution was poured to 1 L of ice water withstirring. The slurry mixture was stirred at room temperature for 2hours. The white precipitate was filtered, washed with water (2×100 mL)and dried under vacuum. The CH₂Cl₂ layer was separated, dried overanhydrous MgSO₄ (50 g), filtered and concentrated under reduced pressureto dryness. The white precipitate and residue from CH₂Cl₂ solution werecombined and purified by flash column chromatography (silica gel,CH₂Cl₂/acetone/MeOH=90/7/3 v/v/v) to give a light tan solid which wasrecrystallized from hot EtOAc/hexanes solution twice to afford a whitecrystalline solid, 104.0 g, 81.6% yield. M.p. 110-112° C. MS (ESI) m/z364.1 [M+Na]⁺, ¹H NMR (DMSO-d₆) δ10.14 (bs, 1H), 9.71 (bs, 1H),7.25-7.11 (m, 5H), 7.05-6.99 (m, 3H), 6.78 (t, J=8.6 Hz, 1H), 6.68 (d,J=8.7 Hz, 2H).

Example 3 Synthesis of the Compound of Formula VI (FIG. 7) Synthesis of4-(benzyloxy)-N-(4-methoxyphenyl)aniline (1d)

A mixture of 4-benzyloxyaniline (16.6 g, 83.31 mmol), 4-iodoanisole(15.0 g, 64.09 mmol), K₂CO₃ (17.72 g, 128.18 mmol), CuI (1.22 g, 6.41mmol) and L-proline (1.48 g, 12.82 mmol) were mixed together anddissolved in anhydrous DMSO (120 mL) at room temperature. Then, thereaction mixture was stirred and heated to 90° C. for 48 hours. Themixture was cooled to room temperature and hydrolyzed with water. EtOAcwas added to partition the solution. The EtOAc layer was separatedwashed with brine, dried over anhydrous MgSO₄. The solvent was removedunder reduced pressure. The solid residue was purified by flash columnchromatography (silica gel) using EtOAc/hexanes (1/9 v/v) to afford thecorresponding diarylaniline as a yellow solid, 9.8 g, 50% yield. M.p.108.0-108.4° C. ¹H NMR (CDCl₃, 300 MHz) δ 7.34-7.25 (m, 5H), 6.90-6.81(m, 8H), 5.02 (s, 2H), 3.78 (s, 3H). MS m/z 306 (M+H)⁺.

Synthesis of N-(4-benzyloxyphenyl)-N-(4-methoxyphenyl)-2-naphthamide(2d)

One equivalent of 4-(benzyloxy)-N-(4-methoxyphenyl)aniline (0.80 g, 2.62mmol) was mixed with 1.5 equivalents of 2-naphthoyl chloride (0.75 g,3.93 mmol) and 4 equivalents of pyridine (0.83 g, 10.48 mmol) in a drythree-necked round-bottomed flask equipped with a magnetic stirring barand a reflux condenser. The mixture was dissolved in anhydrous THF (30mL) and heated to reflux for 20 hours. The reaction solution was cooledto room temperature and filtered. The solvent was removed under reducedpressure. The residue was purified by flash column chromatography usingsilica-gel with EtOAc/hexanes (3/7 v/v) to afford the pure correspondingnaphthamide compound as a white solid, 0.70 g, 58% yield. M.p.174.9-175.5° C. ¹H NMR (CDCl₃, 300 MHz) δ 8.04 (s, 1H), 7.77-7.74 (m,2H), 7.64-7.61 (m, 1H), 7.51-7.43 (m, 4H), 7.40-7.31 (m, 4H), 7.13-7.10(m, 4H), 6.88-6.78 (m, 4H), 4.99 (s, 2H), 3.74 (s, 3H). MS m/z 460(M+H)⁺.

Synthesis of N,N-Bis(4-hydroxyphenyl)-2-naphthylamide (VI)

Compound N-(4-benzyloxyphenyl)-N-(4-methoxyphenyl)-2-naphthamide (2d)(0.50 g, 1.09 mmol) was dissolved in dry CH₂Cl₂ (30 mL) at roomtemperature. BBr₃ (3.26 mL of 1.0 M CH₂Cl₂ solution, 3.26 mmol) wasadded dropwise with stirring via a syringe at room temperature. Thereaction solution was allowed to stir overnight at room temperature. Themixture was cooled to 0° C. in an ice bath and hydrolyzed by addingwater. EtOAc was added to partition the solution. The organic layer wasseparated; the aqueous layer was extracted with EtOAc twice. The organiclayers were combined, washed with brine and dried over anhydrous MgSO₄.The solvent was removed under vacuum. The residue was purified by flashcolumn chromatography using silica-gel with CH₃OH/CH₂Cl₂ (1/9 v/v) toafford the pure desired phenolic compound as a white solid, 0.27 g,white solid, 70% yield. M.p. 264.3-265.2° C. (decomposed). ¹H NMR(DMSO-d₆, 500 MHz) δ 9.46 (s, 2H), 7.98 (s, 1H), 7.85-7.75 (m, 2H),7.75-7.73 (m, 2H), 7.54-7.48 (m, 2H), 7.45-7.43 (m, 1H), 7.05 (s, 4H),6.66 (s, 4H). MS m/z 356 (M+H)⁺.

Example 4 Synthesis of the Compound of Formula VIII

Synthesis of 4-((4-fluorophenyl)(4-hydroxybenzyl)amino)phenol (VIII)

Compound N-(4-fluorophenyl)-4-hydroxy-N-(hydroxyphenyl)benzamide (0.30g, 0.93 mmol) was dissolved in 20 mL anhydrous THF at room temperature.H₃B(SMe₂) (1.86 mL of 2M THF solution, 3.71 mmol) was added via asyringe at room temperature under argon. The reaction solution wasstirred and heated to reflux for 6 hours. Then, the reaction wasquenched by adding 10 mL of MeOH at 0° C. The solvent was removed underreduced pressure. The residue was subjected to flash columnchromatography (silica-gel, CH₂Cl₂/MeOH=9/1 v/v) to give a yellow oil,0.26 g, 92% yield. ¹H NMR (DMSO-d₆, 500 MHz) δ 9.29 (s, 1H), 9.24 (s,1H), 7.09 (d, 2H, J=8.3 Hz), 6.98 (d, 2H, J=9.0 Hz), 6.94-6.91 (m, 2H),6.73 (d, 2H, J=9.0 Hz), 6.68-6.64 (m, 4H), 4.70 (s, 2H). MS m/z 307.8(M−H)⁻.

Example 5 Synthesis of the Compound of Formulas IX and X (FIG. 8)Synthesis of diarylanilines. A mixture of arylamine (1.5 equivalent),aryl iodide (1 equivalent), K₂CO₃ (2 equivalents), CuI (0.1 equivalent)and L-proline (0.2 equivalent) were mixed together and dissolved inanhydrous DMSO at room temperature. Then, the reaction mixture wasstirred and heated to 90° C. for 28 hours. The mixture was cooled toroom temperature and hydrolyzed with water. EtOAc was added to partitionthe solution. The EtOAc layer was separated, washed with brine, driedover anhydrous MgSO₄. The solvent was removed under reduced pressure.The solid residue was purified by flash column chromatography (silicagel) using EtOAc/hexanes (3/7 v/v) as solvent to afford thecorresponding diarylaniline. Bis-(4-methoxyphenyl)amine (1a):pale-yellow solid, 73% yield. ¹H NMR (CDCl₃, 300 MHz) δ 6.93-6.81 (m,8H), 5.37 (s, br, 1H), 3.78 (s, 6H). MS m/z 228.4 (M−H)⁺. Synthesis of4-fluoro-N,N-bis(4-methoxyphenyl)-2-(trifluoromethyl)benzamide (2e)

1 equivalent of bis-(4-methoxyphenyl)amine (1a) (0.73 g, 3.18 mmol) wasmixed with 1.2 equivalents of 4-fluoro-2-trifluoromethylbenzoyl chloride(0.87 g, 3.82 mmol) and 6 equivalents of pyridine (1.51 g, 19.08 mmol)in a dry three-necked round-bottomed flask equipped with a magneticstirring bar and a reflux condenser. The mixture was dissolved inanhydrous THF (20 mL) and heated to 90° C. for 20 hours. The reactionsolution was cooled to room temperature and filtered. The solvent wasremoved under reduced pressure. The residue was purified by flash columnchromatography using silica-gel with EtOAc/hexanes (3/7 v/v) to affordthe pure corresponding benzamide compound as a colorless oil, 1.12 g,84.2% yield. ¹H NMR (CDCl₃, 300 MHz) δ 7.34-7.26 (m, 4H), 7.09-7.01 (m,3H), 6.91 (d, 2H, J=8.7 Hz), 6.87 (d, 2H, J=8.7 Hz), 3.80 (s, 3H), 3.71(s, 3H). MS m/z 442.1 (M+Na)⁺.

Synthesis of4-fluoro-N,N-bis(4-hydroxyphenyl)-2-(trifluoromethyl)benzamide (3a)

Compound 4-fluoro-N,N-bis(4-methoxyphenyl)-2-(trifluoromethyl)benzamide(2e) (1.00 g, 2.38 mmol) was dissolved in dry CH₂Cl₂ (30 mL) at roomtemperature. BBr₃ (10 mL of 1.0 M CH₂Cl₂ solution, 10.0 mmol) was addeddropwise with stirring via a syringe at room temperature. The reactionsolution was allowed to stir overnight at room temperature. The mixturewas cooled to 0° C. in an ice bath and hydrolyzed by adding water. EtOAcwas added to partition the solution. The organic layer was separated;the aqueous layer was extracted with EtOAc twice. The organic layerswere combined, washed with brine and dried over anhydrous MgSO₄. Thesolvent was removed under vacuum. The residue was purified by flashcolumn chromatography using silica-gel with CH₃OH/CH₂Cl₂ (1/9 v/v) toafford the pure desired phenolic compound as a white solid, 0.86 g,92.5% yield. ¹H NMR (DMSO-d₆, 300 MHz) δ 9.55 (s, 1H), 9.53 (s, 1H),7.69-7.58 (m, 2H), 7.46-7.39 (m, 1H), 7.18 (d, 2H, J=8.7 Hz), 6.93 (d,4H, J=8.7 Hz), 7.03 (d, 2H, J=8.4 Hz), 6.78 (d, 2H, J=8.7 Hz), 6.57 (d,2H, J=8.7 Hz). MS m/z 392.1 (M+H)⁺.

Synthesis of4-fluoro-N-(4-hydroxyphenyl)-N-[4-(2-piperidin-1-yl)-ethoxy)phenyl]-2-(trifluoromethyl)benzamide.(IX)

To a solution of4-fluoro-N,N-bis(4-hydroxyphenyl)-2-(trifluoromethyl)benzamide (3a)(0.61 g, 1.56 mmol) in acetone, K₂CO₃ (1.29 g, 9.36 mmol) andN-chloroethyl-piperidine hydrochloride salt (0.34 g, 1.87 mmol) wereadded. The solution was heated to reflux for 20 hours. The solution wasevaporated to dryness. The residue was purified by flash chromatography(silica-gel; methylene chloride/methanol=9/1 v/v) to give the desiredcompound as a white solid, 0.45 g, 57.7% yield. ¹H NMR (DMSO-d₆, 300MHz) δ 9.57 (s, 1H), 7.71-7.68 (m, 2H), 7.47-7.44 (m, 1H), 7.28 (d, 1H,J=9.0 Hz), 7.18 (d, 1H, J=8.7 Hz), 7.13 (d, 1H, J=8.7 Hz), 7.05 (d, 1H,J=8.4 Hz), 6.97 (d, 1H, J=9.0 Hz), 6.80-6.76 (m, 2H), 6.57 (d, 1H, J=8.7Hz), 4.06 (t, 1H, J=6.0 Hz), 3.93 (t, 1H, J=6.0 Hz), 2.66 (t, 1H, J=5.7Hz), 2.55 (t, 1H, J=5.4 Hz), 2.44 (s, 2H), 2.36 (s, 2H), 1.49-1.37 (m,6H). MS m/z 501.0 (M−H)⁻.

The hydrochloride salt (X) was prepared by adding HCl in Et₂O to themethanol solution of the compounds followed by evaporation of solvents.

Example 6 Estrogen Receptor Binding Affinities, Agonist and AntagonistActivity

The ER binding affinity of the compounds was determined using an invitro competitive radioligand binding assay with[2,4,6,7-³H(N)]-Estradiol ([³H]E2), a natural high affinity ER ligand,and bacterially expressed GST fusion ER-α or ER-β ligand binding domain(LBD) protein.

Method

Recombinant ER-α or ER-β was combined with [³H]E₂ to determine theequilibrium dissociation constant (K_(d)) of [³H]E₂. Protein wasincubated with increasing concentrations of [³H]E₂ with and without ahigh concentration of unlabeled E₂ at 4° C. for 18 h in order todetermine total and non-specific binding. Non-specific binding wassubtracted and the K_(d) of E₂ (ERα: 0.71 nM; ERβ: 1.13 nM) wasdetermined using non-linear regression. In addition, the concentrationof [³H]E₂ required to saturate ER-α and ER-β LBD was determined to be4-6 nM.

Increasing concentrations of the compounds (range: 10⁻¹¹ to 10⁻⁶ M) wereincubated with [³H]E₂ (5.7 nM) and ER LBD using the conditions describedabove. Following incubation, plates were harvested with GF/B filters onthe Unifilter-96 Harvester (PerkinElmer) and washed three times withice-cold buffer B (50 mM Tris, pH 7.2). The filter plates were dried atroom temperature, then 35 μl Microscint-O cocktail was added to eachwell and the filter plates were sealed with TopSeal-A. Radioactivity wascounted in a TopCount® NXT Microplate Scintillation Counter using thesettings for ³H in Microscint cocktail (PerkinElmer).

The specific binding of [³H]E₂ at each concentration of the compoundswas determined by subtracting the nonspecific binding of [³H]E₂(determined by incubating with 10⁻⁶ M unlabeled E₂) and expressing it asa percentage of the specific binding in the absence of test compound.The concentration of the compounds that reduced the specific binding of[³H]E₂ by 50% (IC₅₀) was determined. The equilibrium binding constant(K) of the compounds was then calculated by: K_(i)=K_(d)×IC₅₀/(K_(d)+L),where K_(d) is the equilibrium dissociation constant of [³H]E₂(ER-α=0.71 nM; ER-β=1.13 nM), and L is the concentration of [³H]E₂(ER-α: 5.7 nM; ER-β: 5.7 nM).

Results

Binding assays revealed that ligands bound ER-α and ER-β at variousconcentrations ranging from 3.75 nM to greater than 1000 nM andselectivity ranges from the compound being isoform selective to beingnon-isoform selective. Results from representative compounds are listedin Table 2.

TABLE 2 Binding results for selected compounds. ER-α ER-β COMPOUNDSK_(i) (nM) K_(i) (nM)

3.75 81.6

3.81 6.44

21.7 15.2

7.13 35.9

9 72

6.06 76.92

13 19

14.79 646.32

15 57

15.12 25.02

Compound IV binds to ERα and ERβ. The ER binding affinity of Compound IVwas determined using an in vitro competitive radioligand binding assaywith [2,4,6,7-³H(N)]-Estradiol ([³H]E₂), a natural high affinity ERligand, and bacterially expressed GST fusion ERα or ERβ ligand bindingdomain (LBD) protein. In this assay, the ERα and ERβ binding affinities(K_(i) values) of Compound IV were 21.7±1.7 nM (n=3) and 15.2±4.1 nM(n=3), respectively. Upon binding to ER, Compound IV initiates a complexseries of molecular events that lead to the expression or repression oftarget genes involved with pharmacologic response in a tissue-selectivemanner. In transient transfection assays, Compound IV is an ERα and ERβagonist, with greater demonstrated potency to stimulate ERα-mediatedtranscriptional activation as compared to that of ERβ. Whereas estradiolactivates ERα and ERβ with a 5.1-fold greater selectivity for ERα,Compound IV shows a 49.0-fold selectivity for ERα. Thus, Compound IV hasa relative 9.7-fold selectivity in relative transactivation potency(normalized to estradiol values) for ERα over ERβ. Additionally, noantagonist effects were observed in estradiol (1 nM)-stimulatedtranscriptional activation by Compound IV at concentrations up to 10 μM.Although many steroidal ligands cross-react with other nuclear hormonereceptors, the actions of Compound IV are specific for ERα and ERβ.Compound IV was screened for cross-reactivity against rat isoforms ofglucocorticoid receptor (GR), mineralocorticoids receptor (MR),progesterone receptor (PR), androgen receptor (AR) and human isoforms offarnesoid X receptor (FXR), liver X receptor (LXR), peroxisomeproliferator-activated receptors (PPAR-α and PPAR-γ), and retinoid Xreceptor (RXR-α) in both agonist and antagonist modes in transcriptionalactivation assays. Compound IV did not display any agonist or antagonistactivity in any of these assays, supporting the conclusion that CompoundIV does not functionally cross-react with these nuclear hormone receptorsuperfamily members.

Example 7 Transactivation of Selected Compounds

Transactivation assays in agonist and antagonist modes were performed toidentify whether the compound is an agonist, antagonist or a partial.

Method

Rat estrogen receptors (ER-α and ER-β) were cloned from rat ovarian cDNAinto a pCR3.1 plasmid vector backbone. Sequencing was performed todetermine the absence of any mutations. HEK-293 cells were plated at100,000 cells per well of a 24 well plate in Dulbecco's MinimalEssential Media (DMEM)+5% charcoal-stripped fetal bovine serum (csFBS).The cells were transfected using Lipofectamine (Invitrogen, Carlsbad,Calif.) with 0.25 μg ERE-LUC, 0.02 μg CMV-LUC (renilla luciferase) and12.5 ng of rat ER-α or 25 ng rat ER-β. The cells were treated 24 hrsafter transfection with various concentrations of compounds or acombination of compounds and estradiol to determine the antagonisticactivity. Luciferase assays were performed 48 hrs after transfection.

Results

Screening of compounds of this invention in the transactivation systemrevealed that the compounds belonged to all the three classes i.e.agonists, antagonists and partial agonist. An example of an agonist andan antagonist is given in Table-3. Transactivation results matchedextremely well with the binding results for isoform selectivity.

Table 3 provides the EC₅₀ and IC₅₀ transactivation values for someselected compounds of this invention.

TABLE 3 Transactivation (both agonist and antagonist) of selectivecompounds of this invention. ER-α ER-β ER-α ER-β COMPOUND EC₅₀ (nM) EC₅₀(nM) IC₅₀ (nM) IC₅₀ (nM)

    0.65    40.4 >1000 >1000

>1000 >1000     2.207  145

Example 8 Testosterone Suppression in Cynomolgus Monkeys

Two-year old gonadally-intact male Cynomolgus monkeys (n=2) were housedduring the study in compliance with USDA Guidelines with free access toprimate diet and water (except fasted prior to oral doseadministration). Animals were given a once-daily oral gavage dose of 30mg/kg of compound of formula IV in a microemulsion vehicle of Tween80/deionized water for 7 consecutive days. Serum samples were withdrawnby venipuncture prior to the oral dose administration on days 1(baseline), 3, 4, 5, 6, and 7. Testosterone and total androgens werequantified using an enzyme immunoassay (EIA) method combined with orwithout an HPLC method respectively. After 6-days of treatment withcompound of formula IV, time-dependent decreases were apparent fortestosterone and total androgens (testosterone/dihydrotestosterone).Compound of formula IV suppressed the levels of testosterone by 58% and64% in animal #1 and animal #3, respectively, relative to baselinevalues (see solid lines in FIG. 1; Table 4). Similarly, total androgenlevels were suppressed by 56% in both animals #1 and #3 (see dashedlines in FIG. 1; Table 4) compared to baseline values.

Consistent with estrogen feedback of the pituitary-testicular axis inmales, these results demonstrate a robust pharmacologic response for thesuppression of serum hormones (testosterone and total androgens) inintact non human primates (Cynomolgus monkeys) after repeated oral doses(30 mg/kg) of compound of formula IV.

TABLE 4 Testosterone and total androgen levels in serum of intact malemonkeys with daily 30 mg/kg oral administration compound of formula IV(first dose on Day 0). Day Animal 1 Animal 3 Animal 1 Animal 3Testosterone Total Androgens Serum level (pg/mL) Serum level (pg/mL) 01120 617 1868 1643 (baseline) 2 937 479 1178 847 3 784 437 1078 786 4552 415 988 924 5 403 276 966 664 6 474 221 819 726 Percent reductionfrom Percent reduction from baseline baseline 0 100 100 100 100(baseline) 2 16 22 37 48 3 30 29 42 52 4 51 33 47 44 5 64 55 48 60 6 5864 56 56

Example 9

Suppression of LH and Testosterone Hormone Levels in Rats

An in vivo dose-response study was conducted to evaluate the effect ofCompound IV on LH suppression in intact and orchiectomized (ORX) malerats. In intact and ORX animals, Compound IV at doses ≧10 mg/kg per daysignificantly suppressed LH levels when compared to respective controls.(The same pattern of suppression was observed in FSH levels.) LHsuppression resulted in robustly decreased testosterone levels to belowthe limit of quantitation (BLOQ) which is 0.08 ng/mL and decreasedweights of prostate, seminal vesicles, and levator ani weights musclesince these are highly androgen-dependent organs. In intact animals,dose-dependent decreases in the weights of these target organs werenoted with the seminal vesicles and levator ani muscle weights to thelevel of castrated controls. Although prostate weights weresignificantly reduced in intact animals, these values did not reach thelevel of castrated controls. Results are summarized in Table 6hereinbelow.

Materials and Methods:

Male Sprague-Dawley rats weighing approximately 200 g were maintained ona 12-h light/dark cycle with food (2016 Teklad Global 16% Protein RodentDiet, Harlan, Madison, Wis.) and water available ad libitum. The animalprotocol was reviewed and approved by the Institutional Animal Care andUse Committee of the University of Tennessee.

The test article for this study was weighed and dissolved in 10% DMSO(Fisher) diluted with PEG 300 (Acros Organics, NJ) to prepare theappropriate dose formulations. For this study, sixty (60) maleSprague-Dawley rats were randomized by body weight, and assigned to oneof the twelve treatment groups (n=5 animals/group). Treatment groups arelisted in Table 5. The animals were housed in groups of 2 to 3 animalsper cage. Control groups (intact and orchidectomized (ORX)) wereadministered vehicle daily. Compound IV was administered viasubcutaneous injection (200 μL) at doses of 0.3, 1, 3, 10, and 30mg/kg/day to both intact and ORX groups.

After a 14-day dosing regimen, the animals were sacrificed underanesthesia (ketamine/xylazine, 87:13 mg/kg) and body weights wererecorded. In addition, ventral prostate, seminal vesicles, and levatorani muscle were removed, cleaned of extraneous tissue, and individuallyweighed. Organ weight were normalized to body weight and expressed as apercentage of intact control. Blood was collected from the abdominalaorta under isoflurane anesthesia and allowed to clot. Serum wasseparated by centrifugation and stored at −80° C. prior to determinationof serum hormone levels. Serum luteinizing hormone (LH) and folliclestimulating hormone (FSH) concentrations were determined by the RatPituitary Luminex Assay (Millipore, Billerica, Mass.) according tomanufacturer's directions. The lower limit of quantitation for thisassay was 3.2 pg/mL for LH and 32 pg/mL for FSH. Testosterone wasmeasured by a Testosterone EIA (Alpco Diagnostics, Salem, N.H.) with aLLOQ of 0.08 ng/mL. Serum hormone values below the lower limit ofquantitation (BLOQ) were omitted from analysis of group means.Therefore, the reported value for LH and T in the groups with samplesBLOQ is higher than the actual value. This method of analysis providedthe most conservative estimate of LH and T suppression. Fisher's LeastSignificant Difference test was used to compare individual dose groupsto the intact and ORX vehicle control groups. Significance was defined apriori as a P-value <0.05.

TABLE 5 Treatment groups. Group Gonadal Status Dose (mg/kg/day) TestArticle 1 Intact — Vehicle 2 ORX — Vehicle 3 Intact 0.3 Compound IV 4Intact 1 Compound IV 5 Intact 3 Compound IV 6 Intact 10 Compound IV 7Intact 30 Compound IV 8 ORX 0.3 Compound IV 9 ORX 1 Compound IV 10 ORX 3Compound IV 11 ORX 10 Compound IV 12 ORX 30 Compound IV

Luteinizing Hormone Levels in Intact and ORX Rats (Table 6)

LH levels (mean±SD) in intact and ORX vehicle control groups were1.46±0.64 and 11.1±3.9 ng/mL, respectively. Compound IV dose-dependentlyreduced LH levels in intact animals, reaching statistically significantreductions with daily doses ≧3 mg/kg. LH levels in intact Compound IVtreated animals were 0.863±0.384, 0.704±0.530, 0.395±0.302, 0.226±0.165,and 0.236±0.176 ng/mL, following doses of 0.3, 1, 3, 10, and 30mg/kg/day, respectively. LH levels in ORX males were also significantlydecreased by Compound IV treatment. In ORX animals the LH levels were15.4±2.9, 13.5±2.2, 6.5±5.6, 0.425±0.135, and 0.368±0.119 ng/mL,following doses of 0.3, 1, 3, 10, and 30 mg/kg/d, respectively. Theresults are presented graphically in FIG. 10A.

Follicle Stimulating Hormone Levels in Intact and ORX Rats (Table 6)

Serum FSH levels in intact and ORX vehicle control groups were 20.9±8.5and 93.5±13.8 ng/mL, respectively. In intact animals, Compound IVdose-dependently reduced FSH levels with significant reductions observedat doses ≧10 mg/kg/day. FSH levels in intact Compound IV treated animalswere 17.3±6.4, 15.7±7.3, 18.4±7.7, 9.2±4.0, and 6.3±1.8 ng/mL, followingdoses of 0.3, 1, 3, 10, and 30 mg/kg/day, respectively. In ORX animalsthe LH levels were 115±17, 114±22, 65.2±31.9, 27.6±8.2, and 15.1±4.1ng/mL, following doses of 0.3, 1, 3, 10, and 30 mg/kg/day, respectively.The results are presented graphically in FIG. 10B.

Testosterone Levels in Intact and ORX Rats

Serum testosterone levels in intact vehicle control groups were 2.4±1.1ng/mL. The lower limit of quantitation for T was 0.08 ng/mL. Values lessthan 0.08 ng/mL are designated as Below the Limit Of Quantitation(BLOQ). In intact animals, compound of formula IV dose-dependentlyreduced T levels with significant reductions observed at doses ≧3 mg/kgper day. Testosterone levels in intact animals treated with compound offormula IV were 2.6±1.7, 1.6±1.0, 0.7±0.4, BLOQ, and BLOQ ng/mL,following doses of 0.3, 1, 3, 10, and 30 mg/kg per day, respectively. InORX animals the T levels were BLOQ for all groups treated with compoundIV and the vehicle treated group. The results are for the intact animalsare presented graphically in FIG. 10C (and FIG. 2) (BLOQ values arerepresented at the limit of quantitation for graphical purposes).

Rapid and potent suppression of serum testosterone in intact male ratswas measured by administering Compound IV with dosages of 3 mg/kg, 10mg/kg and 300 mg/kg after 24 h, 72 h and 168 h as presented in FIG. 9.

Organ Weights (Table 6)

Prostate, seminal vesicles, and levator ani muscle weights were measuredto confirm the suppression of T. The organ weights (mean±SD) arepresented in FIGS. 10D, 10E and 10F respectfully. Dose-dependantdecreases in prostate, seminal vesicles, and levator ani muscle weightwere observed in intact animals treated with Compound IV. Prostateweights in intact animals were 84.0±19.2, 75.2±20.7, 68.2±8.1,45.1±20.0, and 43.6±8.8, following doses of 0.3, 1, 3, 10, and 30mg/kg/day, respectively. Prostate weights in ORX animals were 19.0±4.2,17.4±3.4, 19.6±6.7, 22.9±5.4, and 20.6±2.1, following doses of 0.3, 1,3, 10, and 30 mg/kg/day, respectively. Seminal vesicle weights in intactanimals were 76.2±7.8, 66.3±27.2, 51.8±28.5, 19.1±7.0, and 17.9±3.3,following doses of 0.3, 1, 3, 10, and 30 mg/kg/day, respectively.Seminal vesicle weights in ORX animals were 12.2±1.3, 16.6±5.4,16.5±4.8, 13.3±1.9, and 12.9±2.1, following doses of 0.3, 1, 3, 10, and30 mg/kg/day, respectively. Levator ani weights in intact animals were86.9±10.0, 82.1±12.1, 65.2±4.4, 57.8±11.2, and 58.1±4.7, following dosesof 0.3, 1, 3, 10, and 30 mg/kg/day, respectively. Levator ani weights inORX animals were 54.5±6.6, 49.6±7.0, 53.6±10.0, 51.1±4.9, and 49.2±4.2,following doses of 0.3, 1, 3, 10, and 30 mg/kg/day, respectively.

The LH suppression and organ weights data are summarized in Table 6.

TABLE 6 In vivo effects of the compound of formula IV on serum hormonesand organ weight. Dose Seminal Levator Ani Gonadal (mg/kg LH FSHProstate Vesicles Muscle Status Compound per day) (ng/mL) (ng/mL) (% ofIntact) (% of Intact) (% of Intact) Intact Vehicle — Mean 1.46^(b)20.9^(b)   100.0^(b) 100.0^(b) 100.0^(b) S.D. 0.642 8.49 28.6 13.4 4.97ORX Vehicle — Mean 11.1^(a) 93.5^(a)   13.7^(a) 14.0^(a) 58.8^(a) S.D.3.87 13.8  2.56 2.93 6.62 Intact Compound 0.3 Mean 0.863^(b) 17.3^(b)  84.0^(b) 76.2^(a,b) 86.9^(a,b) IV S.D. 0.384 6.44 19.2 7.83 10 IntactCompound 1 Mean 0.704^(b) 15.7^(b)   75.2^(b) 66.3^(a,b) 82.1^(a,b) IVS.D. 0.53 7.26 20.7 27.2 12.1 Intact Compound 3 Mean 0.395^(a,b)18.4^(b)   68.2^(a,b) 51.8^(a,b) 65.2^(a) IV S.D. 0.302 7.72 8.12 28.54.35 Intact Compound 10 Mean 0.226^(a,b)   9.25^(a,b) 45.1^(a,b)19.1^(a) 57.8^(a) IV S.D. 0.165 3.97 20 6.98 11.2 Intact Compound 30Mean 0.236^(a,b)   6.25^(a,b) 43.6^(a,b) 17.9^(a) 58.1^(a) IV S.D. 0.1761.82 8.75 3.33 4.71 ORX Compound 0.3 Mean 15.4^(a) 116^(a)   19.0^(a,b)12.2^(a) 54.5^(a) IV S.D. 2.94 17.2  4.19 1.31 6.56 ORX Compound 1 Mean13.5^(a) 114^(a)   17.4^(a) 16.6^(a) 49.6^(a) IV S.D. 2.18 22.3  3.45.36 7.04 ORX Compound 3 Mean 6.5 65.2^(a)   19.6^(a) 16.5^(a) 53.6^(a)IV S.D. 5.63 31.9  6.67 4.82 10 ORX Compound 10 Mean 0.425^(a,b)27.6^(b)   22.9^(a,b) 13.3^(a) 51.1^(a) IV S.D. 0.135 8.16 5.44 1.914.88 ORX Compound 30 Mean 0.368^(a,b) 15.1^(b)   20.6^(a,b) 12.9^(a)49.2^(a,b) IV S.D. 0.119 4.11 2.08 2.14 4.21 ^(a)P < 0.05 versus IntactVehicle. ^(b)P < 0.05 versus ORX Vehicle

Example 10 Recovery of Testosterone Levels Following Suppression byCompound IV in Rats and Monkeys

The reversibility of chemical castration with Compound IV was studied.

Materials and Methods:

Thirty-five (35) male Sprague-Dawley rats weighing approximately 200 gwere maintained on a 12-h light/dark cycle with food (2016 Teklad Global16% Protein Rodent Diet, Harlan, Madison, Wis.) and water available adlibitum. The animal protocol was reviewed and approved by theInstitutional Animal Care and Use Committee of the University ofTennessee.

The test article for this study was weighed and dissolved in PEG 300(100%) (Acros Organics, NJ) to prepare the appropriate doseformulations. Animals were randomly assigned to one of the ten treatmentgroups (n=5 animals/group). Treatment groups are listed in Table 7. Theanimals were housed in groups of 2 to 3 animals per cage. Group 1 wassacrificed at the initiation of the study (Day 1) for determination ofbaseline testosterone levels in intact animals. Groups 2-7 receiveddaily doses of 1, 3, or 30 mg/kg via oral gavage (˜200 uL) for threedays. Groups 2, 3, and 4 were sacrificed on Day 4 to measure maximaltestosterone suppression. Groups 5, 6, and 7 were allowed to recover for14 days with a drug free washout period.

TABLE 7 Treatment groups. Compound IV Group P.O. Dose Treatment Group 1— Baseline Group 2  1 mg/kg for 3 days No Recovery Group 3  3 mg/kg for3 days No Recovery Group 4 30 mg/kg for 3 days No Recovery Group 5  1mg/kg for 3 days 14 day recovery Group 6  3 mg/kg for 3 days 14 dayrecovery Group 7 30 mg/kg for 3 days 14 day recovery

Results:

Serum testosterone levels in intact rats were 6.4±3.1 ng/mL (mean±S.D)at baseline. Compound IV administered at doses of 3 and 30 mg/kg forthree days significantly suppressed serum testosterone levels to1.47±0.26 and 1.62±0.49 ng/mL, respectively. No significant suppressionwas observed in animals that received 1 mg/kg of Compound IV for threedays. Most importantly, serum testosterone levels were 3.3±1.92,3.00±1.06 and 3.8±1.72 in animals that received 1, 3, or 30 mg/kg,respectively, of Compound IV for three days when measured after a 14 dayrecovery period, and were not statistically significantly differencesfrom baseline serum testosterone concentrations in intact rats asdepicted in FIG. 23.

This study confirms previous results showing that Compound IV quicklysuppresses serum testosterone levels in intact male rats. We observedsuppression of serum testosterone levels in dose groups receiving ≧3mg/kg/day for 3 days. A significant decrease in serum testosterone wasnot observed with the 1 mg/kg dose group. However, within 14 days ofrecovery, serum testosterone levels had returned back to the level ofintact controls. This study shows that pharmacologic castration byCompound IV is reversible in rats.

The effect of Compound IV on suppression and recovery of testosteronelevels in intact male monkeys was evaluated in conjunction with an oralpharmacokinetic study. Three treatment naïve male Cynomolgus monkeys (2to 3 years old) were administered Compound IV at 30 mg/kg daily by oralgavage for 7 consecutive days. Blood samples were collected and dividedinto serum and plasma for testosterone and Compound IV quantitativemeasurements, respectively. Results show that daily oral doses ofCompound IV significantly decreased circulating androgen (primarilytestosterone and dihydrotestosterone) levels in all three male monkeysby up to 47% compared to baseline levels (levels of 1591±72.5, 997±104,and 852±136 ng/mL, respectively for baseline, Day 2 and Day 6 oftreatment [mean±SEM]). Following a 18-day drug-free recovery period,androgen levels returned to normal, and were not significantly differentfrom pre-treatment baseline levels (1757.7±369.5 ng/mL after recovery).

Example 11 Bone Preservation Despite Reduction of LH and Testosterone inRats Table 8

The effect of compound of formula IV on treatment on bone was studied.Orally administered compound of formula IV completely prevented the boneloss associated with LH suppression in intact male rats. Significantreduction of LH was induced by the compound of formula IV in intactanimals at dose levels ≧10 mg/kg per day. Although at 1 mg/kg per day,compound of formula IV did not significantly reduce LH, significantreductions in prostate, seminal vesicles, and levator ani muscle wereapparent at this dose indicating that the reduction in circulatingtestosterone was physiologically relevant to these androgen responsiveorgans. However, 1 mg/kg per day compound of formula IV maintainedtrabecular bone volume (measured in the distal femur) at the level ofintact controls. When administered at doses of 10 and 30 mg/kg per day,compound of formula IV increased bone volume in the distal femursignificantly above that of intact controls. These data show thatcompound IV increased trabecular bone mineral density (BMD) and percentbone volume at a dose level that reduces LH levels in intact rats. Datafrom this study are presented in Table 8.

TABLE 8 In vivo effects of Compound IV on rat bone, organ weight, andserum hormone parameters. Bone Dose Mineral Percent Seminal Levator AniGonadal (mg/kg Density Bone Volume Prostate Vesicles Muscle FSH LHStatus Compound per day) (g/cm³) (BV/TV) (%) (% of Intact) (% of Intact)(% of Intact) (ng/mL) (ng/mL) Intact Vehicle — Mean 0.274^(b) 20.2^(b)100.0^(b) 100.0^(b) 100.0^(b)  9.93^(b) 0.781^(b) S.D. 0.033 3.57 11.115.9 11.6 2.94 0.263 ORX Vehicle — Mean 0.224^(a) 15.4^(a) 14.8^(a)10.3^(a) 59.4^(a) 117^(a)   22.0^(a) S.D. 0.025 2.6 4.08 0.767 7.2640.2  5.81 Intact Compound  1 Mean 0.273^(b) 20.0^(b) 69.2^(a,b)44.6^(a,b) 80.0^(a,b) 14.1^(a,b) 0.820^(b) IV S.D. 0.04 4.08 13.5 15.76.69 4.07 0.392 Intact Compound 10 Mean 0.326^(a,b) 25.9^(a,b)30.7^(a,b) 12.8^(a,b) 58.1^(a)   5.48^(a,b) 0.060^(a,b) IV S.D. 0.0484.76 12.4 0.886 9.68 1.97 0.092 Intact Compound 30 Mean 0.326^(a,b)25.5^(a,b) 30.1^(a,b) 14.4^(a,b) 56.1^(a)   6.32^(a,b) 0.078^(a,b) IVS.D. 0.046 4.49 17.4 1.45 4.67 3.4  0.114 ^(a)P < 0.05 versus IntactVehicle. ^(b)P < 0.05 versus ORX Vehicle

Example 12 Effects on 17β Hydroxysteroid Dehydrogenase 5 (17β-HSD5)Enzyme Activity

HSD family members are involved in the conversion of circulatingsteroids. 17β-HSD5 converts androstenedione to testosterone and estroneto estradiol. In addition, it is also involved in prostaglandinsynthesis. Here the ability of some select compounds of this inventionto inhibit 17β-HSD5 activity was demonstrated.

Method

Human 17β-HSD5 was cloned in pGEX 4t1 vector and purified protein wasprepared. The purified protein was incubated with the representativecompound of this invention, ¹⁴C androstenedione and NADPH in anappropriate buffer. The synthesized testosterone was extracted usingethyl acetate, air dried, spotted and run on a thin layer chromatography(TLC) plate. The TLC was exposed to phosphorimager and the intensity oftestosterone band was quantified. Indomethcin was used as a positivecontrol (LHRH agonist).

Results

Compound IV was tested and had partial inhibitory effect on 17β-HSD5enzyme activity. The positive control (LHRH agonist), indomethacin asexpected exhibited strong inhibition of this enzyme, as presented inFIG. 3.

Example 13 Toxicity Studies

A study was conducted to compare the thrombotic potential of Compound IVand diethylstilbestrol (DES, positive control) using the in vitro humanplatelet aggregation assay. Blood from healthy male donors was used inthe study since males are the intended treatment population for CompoundIV (LH suppression). Platelet rich plasma was preincubated with eitherestradiol (E2), DES, Compound IV or vehicle for 30 seconds, and thenthrombin (0.3 units) was added to induce platelet aggregation. Resultsof the study show that preincubation with DES increased thethrombin-induced platelet aggregation by approximately 10-fold. However,Compound IV and estradiol decreased aggregation in the platelet richplasma. These data demonstrate that Compound IV reduced the reactivityof human platelets in vitro compared to DES, and suggest that CompoundIV may have lower thromboembolic potential than DES (FIG. 4).

Example 14 Effect of Compound IV on Hot Flashes

A study was conducted to investigate the effect of Compound IV on hotflashes using the morphine dependent rat model (MD model) which wasdeveloped by Simpkins et. al (1983) and was shown to have severalsimilarities to the menopausal hot flush. In addition to thesimilarities to the human condition, this experimental animal model hasa short turn around time which makes it a useful high throughputscreening tool for identifying compounds that can alleviate vasomotorsymptoms using the tail skin temperature (TST). TST probes TA-40 (DataSciences International, MN) were taped to the base of the tails andbaseline temperatures were obtained for 15 minutes. After 15 minutes theanimals were treated with naloxone (1 mg/kg, SQ) to reverse the effectsof the morphine. Tail skin temperature (TST) was measured for one hourpost-naloxone treatment with a sampling frequency of 5 secs throughoutthe course of the experiment. Following the data acquisition, the movingaverage of the temperature recorded every 60 seconds for each animal wascalculated and further analyzed. Baseline temperature was computed asthe average temperature acquired over the 15 minutes preceding naloxoneadministration. The area under the curve (AUC) was calculated bysubtracting all the values post-naloxone administration from thebaseline using a linear trapezoid method.

Compound IV attenuated hot flushes in the morphine withdrawal model (seeFIG. 13) with the best results at 10 mg Compound IV. 17β E2 was used at5 mg/kg in 100% DMSO.

Example 15 Compound IV Versus DES in Rats

Prior to the introduction of LHRH agonists, castrate testosterone levelswere achieved by increasing estrogen activity in the pituitary viaestrogens, primarily diethylstilbestrol (DES). DES was equally effectiveas LHRH agonists at suppressing testosterone to castrate levels.Patients treated with DES did not have hot flashes or bone loss, but didhave gynecomastia at higher rates than ADT with LHRH agonists.Unfortunately, highly potent, pure estrogens, like DES and estradiol,are often associated with a high risk of severe cardiovascular andthromboembolic complications which have limited their clinical use. Ithas been hypothesized, but not proven, that the increased risk of venousthromboembolic complications with DES is due to its cross-reactivitywith other hormone receptors. In vitro studies with human plateletsshowed that Compound IV had much lower procoagulatory activity than DES.Thus, Compound IV, an ER-alpha selective agonist, may deliver theprostate cancer benefits of DES and also deliver the benefits of an LHRHagonist without causing osteoporosis or adverse lipid profiles.

Compound IV is as effective as DES in reducing prostate size in rats(FIG. 11A) and presenting moderate increase in prostate size of ORX rats(FIG. 11B).

Differences between DES and Compound IV are presented in FIGS. 12A-12C,where DES crossreacted with glucocorticoid receptor (GR) (FIG. 12A) andandrogen receptor (AR) (FIG. 12B) while Compound IV did not. Inaddition, DES antagonized estrogen related receptor (ERR)transactivation while Compound IV did not. Compound IV failed tocrossreact with any of the three ERR isoforms (ERR-β, ERR-β and ERR-γ)as depicted in FIG. 12C.

Example 16 Monkey Toxicity Study 90 Days

Colony-bred cynomolgus macaques of Mauritius origin were obtained. Theprospective study was designed as a 39-week oral pharmacology andtoxicology evaluation of Compound IV and positive control (LHRH agonist)in the male cynomolgus monkey with a 13-week interim period. A total of39 sexually mature male monkeys, 5 to 8 years of age, were randomlyassigned to five groups prior to treatment initiation. Groupsincluded: 1) vehicle control, 2) 1 mg/kg Compound IV, 3) 10 mg/kgCompound IV, 4) 100 mg/kg Compound IV, and 5) positive control (LHRHagonist). Drug was delivered orally by cage-side administration oncedaily for 39 weeks with vehicle control article (Tween 80/PRANG™) forGroups 1 and 5, or Compound IV in vehicle for Groups 2, 3, and 4. Doselevels of Compound IV were 1, 10, and 100 mg/kg/day for Groups 2, 3, and4, respectively. Oral doses were delivered in a 10 mL/kg dose volume ascalculated based on most recent available body weight for each animal(FIG. 14). Animals in Group 5 also received a once-daily subcutaneousinjection of positive control (LHRH agonist) (0.02 mL constant volume)for the 39 week study period. General appearance and clinical signs wereobserved and recorded daily. Routine evaluations and select other studyinvestigations were performed as indicated in the study protocol. Selectparameters include, but are not limited to, testosterone, prostatespecific antigen (PSA), and prostate volume and weight.

Testosterone and total PSA levels were quantified in serum samples(following standard procedure) using an enzyme immunoassay (EIA) methodand chemiluminescence immunoassay (LIA, ALPCO Diagnostics, Salem N.H.),respectively. Blood samples for testosterone evaluations were collectedfrom all animals (in fasted state) at baseline (i.e., prior tocommencement of treatment) and on Days 1, 3, 7, 14, 28, 64, and 90.Blood samples for PSA determinations were collected from all animals (infasted state) at baseline and during Week 6.

For the purpose of discussion, results for samples with concentrationsbelow the limit of quantitation (BLQ) for the testosterone and PSAassays are calculated as ½ of the lower limit of quantitation (LLOQ) ofthe assay, and are considered as “Estimated final concentrations”. Datain Tables 9 through 16 are presented as “Quantifiable concentrationsonly” (i.e., excludes BLQ values) in addition to “Estimated finalconcentrations” (i.e., samples with BLQ result included as ½ LLOQ ofassay). Prostate volume was measured in live animals under anesthesiausing a transrectal ultrasound (TRUS) procedure at baseline and Week 6.The width and height of prostate were recorded. Prostate volumes werecalculated as width×width×height×pi/6 and were normalized to bodyweight. The wet weight of prostate was recorded at necropsy aftertrimming the tissue free of fat and extraneous tissue.

Results and Discussion:

Serum testosterone levels are presented in FIG. 15 and Tables 9 through12. At baseline, the testosterone levels for all monkeys on the studywere in the normal range for sexually mature adult male cynomolgusmonkeys. However, testosterone levels were significantly reduced inmonkeys receiving Compound IV at 100 mg/kg/day and in monkeys treatedwith positive control (LHRH agonist). Testosterone levels in thepositive control (LHRH agonist) group illustrated a biphasic change,with an initial significant increase (i.e., flare) of 47.4% and 54.7%(p<0.01) on Days 1 and 3, respectively, followed by decreases of 3.6%,67%, 73%, 83%, and 85% on Days 7, 14, 28, 64 and 90 (see FIG. 15 andTables 9 to 12). A similar flare was not observed for any animal treatedwith Compound IV even at the highest dose level (i.e., 100 mg/kg/day).Dose and treatment duration were important to the pharmacologic actionof Compound IV, where doses of 100 mg/kg/day suppressed the serumtestosterone by 60%, 51%, 42%, 79% and 92% on Days 3, 7, 14, 28 and 64,respectively, relative to the baseline value (see FIG. 15 and Tables 9and 10). After 90 days of treatment with 100 mg/kg/day Compound IV, thetestosterone level in 6 of 10 Group 4 monkeys was reduced toconcentrations below the limit of quantification of the assay (refer toTable 11). The mean serum testosterone level in Group 4 monkeys wasreduced by 96% compared to respective baseline values (“Estimated finalconcentrations”, i.e., testosterone levels for 6/10 monkeys with BLQvalues are calculated as 50% of the LLOQ concentration, see Table 10).It is important to note that by Day 90, Compound IV at 100 mg/kg/dayreduced serum testosterone to levels significantly lower than thepositive control (LHRH agonist) (p=0.013).

TABLE 9 Mean serum testosterone levels (ng/mL) in intact male monkeysafter daily oral administration of Compound IV; ^(@)Estimated finalconcentrations. Vehicle Compound IV Compound IV Compound IV Positivecontrol Control 1 mg/kg 10 mg/kg 100 mg/kg (LHRH agonist) Day Mean SEM NMean SEM N Mean SEM N Mean SEM N Mean SEM N 0 baseline 6.1 1.2 10 7.31.0 6 4.9 0.6 6 4.4 0.6 10 4.9 0.9 7 1 8.0 1.7 10 11 1.6 6 7.6 1.1 6 8.02.2 10 7.2 0.8 7 3 8.2 2.3 10 7.4 1.2 6 5.1 1.1 6 1.8* 0.5 10 32^(#$) 3.8 7 7 5.9 1.2 10 6.7 0.8 6 7.7 1.9 6 2.2* 0.7 9 4.7 2.6 7 14 3.4 0.510 3.8 0.4 6 7.1 1.6 6 2.6 0.9 9  1.6^(#) 0.2 7 28 3.8 0.6 10 4.7 0.9 69.4 2.1 6 0.9* 0.2 10  1.3^(#) 0.2 7 64 5.1 1.1 10 4.3 0.6 6 5.4 1.5 60.3* 0.1 9   0.8^(#$) 0.2 7 90 3.6 0.6 9 4.2 0.6 4 4.6 1.0 5 0.2* 0.0 10  0.8^(#$) 0.2 7 Testosterone assay LLOQ = 0.246 ng/mL; ^(@)BLOQ valuesare calculated as 0.123 ng/mL, half of the LLOQ. *Statisticallysignificant (p < 0.05) Compound IV 100 mg/kg vs. Vehicle Control^(#)Statistically significant (p < 0.05) Positive control (LHRH agonist)vs. Vehicle Control ^($)Statistically significant (p < 0.05) Positivecontrol (LHRH agonist) vs. Compound IV 100 mg/kg

TABLE 10 Percentage change (%) of mean serum testosterone levelscompared to baseline; ^(@)Estimated final concentrations. CompoundVehicle Compound IV Compound IV IV Positive Day Control 1 mg/kg 10 mg/kg100 mg/kg Control 1 31 44 54 82 47 3 35 1.8 3.5 −60 547 7 −3.2 −8.1 57−51 −3.6 14 −44 −48 45 −42 −67 28 −38 −35 92 −79 −73 64 −16 −41 11 −92−83 90 −42 −42 −5.5 −96 −85 Testosterone assay LLOQ = 0.246 ng/mL;^(@)BLQ values are calculated as 0.123 ng/mL, half of the LLOQ.

TABLE 11 Mean serum testosterone levels (ng/mL) in intact male monkeysafter daily oral administration Compound IV; ^(λ)Quantifiableconcentrations only Vehicle Compound IV Compound IV Compound IV Positivecontrol Control 1 mg/kg 10 mg/kg 100 mg/kg (LHRH agonist) Day Mean SEM NMean SEM N Mean SEM N Mean SEM N Mean SEM N 0 baseline 6.1 1.2 10 7.31.0 6 4.9 0.6 6 4.4 0.6 10 4.9 0.9 7 1 8.0 1.7 10 11 1.6 6 7.6 1.1 6 8.02.2 10 7.2 0.8 7 3 8.2 2.3 10 7.4 1.2 6 5.1 1.1 6 1.8 0.5 10 32 3.8 7 75.9 1.2 10 6.7 0.8 6 7.7 1.9 6 2.2 0.7 9 4.7 2.6 7 14 3.4 0.5 10 3.8 0.46 7.1 1.6 6 2.6 0.9 9 1.6 0.2 7 28 3.8 0.6 10 4.7 0.9 6 9.4 2.1 6 0.90.2 10 1.3 0.2 7 64 5.1 1.1 10 4.3 0.6 6 5.4 1.5 6 0.3 0.1 9 0.8 0.2 790 3.6 0.6 9 4.2 0.6 4 4.6 1.0 5 0.2 0.1 4 0.8 0.2 7 Testosterone assayLLOQ = 0.246 ng/mL; ^(λ)BLQ values are excluded.

TABLE 12 Percentage change (%) of mean testosterone levels compared tobaseline; ^(λ)Quantifiable concentrations only. Compound VehicleCompound IV Compound IV IV Positive Day Control 1 mg/kg 10 mg/kg 100mg/kg Control 1 31 44 54 82 47 3 35 1.8 3.5 −60 547 7 −3.2 −8.1 57 −51−3.6 14 −44 −48 45 −42 −67 28 −38 −35 92 −79 −73 64 −16 −41 11 −92 −8390 −42 −42 −5.5 −95 −85 Testosterone assay LLOQ = 0.246 ng/mL; ^(λ)BLQvalues are excluded.

Serum PSA levels were also significantly suppressed by Compound IVwithin four weeks of treatment initiation. PSA reductions of 69% and 87%(in mean) were noted for monkeys receiving Compound IV at 10 mg/kg and100 mg/kg for 4 weeks, whereas PSA levels were reduced by 60% in thepositive control (LHRH agonist) group (FIG. 16 and Tables 13-16).

TABLE 13 Mean serum PSA levels (ng/mL) in intact male monkeys afterdaily oral administration of Compound IV; ^(@)Estimated finalconcentrations. Vehicle Compound IV Compound IV Compound IV Positivecontrol Control 1 mg/kg 10 mg/kg 100 mg/kg (LHRH agonist) Mean SEM NMean SEM N Mean SEM N Mean SEM N Mean SEM N Pre-dose 1.1 0.2 10 1.0 0.26 0.8 0.1 6 1.0 0.1 10 1.0 0.1 7 4-week 1.0 0.2 10 0.9 0.2 6 0.3* 0.1 60.1^(&) 0.1 10 0.4^(#$) 0.1 7 PSA assay LLOQ = 0.0575 ng/mL; ^(@)BLQvalues are calculated as 0.02875 ng/mL, half of the LLOQ. *Statisticallysignificant (p < 0.05) Compound IV 10 mg/kg vs. Vehicle Control^(&)Statistically significant (p < 0.05) Compound IV 100 mg/kg vs.Vehicle Control ^(#)Statistically significant (p < 0.05) Positivecontrol (LHRH agonist) vs. Vehicle Control ^($)Statistically significant(p < 0.05) Positive control (LHRH agonist) vs. Compound IV 100 mg/kg

TABLE 14 Percentage change (%) of mean PSA levels compared to baseline;^(@)Estimated final concentrations. Positive Compound Compound controlIV Compound IV IV (LHRH Control 1 mg/kg 10 mg/kg 100 mg/kg agonist)4-week −7.1 −11 −69 −87 −60 PSA assay LLOQ = 0.0575 ng/mL; ^(@)BLQvalues are calculated as 0.02875 ng/mL, half of the LLOQ.

TABLE 15 Mean serum PSA levels (ng/mL) in intact male monkeys afterdaily oral administration Compound IV; ^(λ)Quantifiable concentrationsonly. Vehicle Compound IV Compound IV Compound IV Positive controlControl 1 mg/kg 10 mg/kg 100 mg/kg (LHRH agonist) Mean SEM N Mean SEM NMean SEM N Mean SEM N Mean SEM N Pre-dose 1.2 0.2 9 1.0 0.2 6 0.8 0.1 61.0 0.1 10 1.0 0.1 7 4-week 1.1 0.1 9 0.9 0.2 6 0.3 0.1 5 0.3 0.1 4 0.40.1 7 PSA assay LLOQ = 0.0575 ng/mL; ^(λ)BLQ values are excluded in thistable.

TABLE 16 Percentage change (%) of mean PSA levels compared to baseline;^(λ)Quantifiable concentrations only. Compound Compound IV Compound IVIV Positive Control 1 mg/kg 10 mg/kg 100 mg/kg Control 4-week −7.1 −11−64 −72 −60 PSA assay LLOQ = 0.0575 ng/mL; ^(λ)BLQ values are excludedin this table.

Prostate volumes were measured by TRUS periodically throughout thestudy. Results obtained after six weeks of treatment demonstrate apotent effect of Compound IV and positive control (LHRH agonist) onmonkey prostate. Compound IV significantly suppressed prostate volumesby 25% and 45% at the 10 mg/kg and 100 mg/kg dose levels, respectively,whereas prostate volumes were reduced by 28% in the positive control(LHRH agonist) group (FIG. 17 and Tables 17 and 18).

TABLE 17 Mean prostate volumes (ratio) in male monkeys after daily oraladministration Compound IV. Vehicle Compound IV Compound IV Compound IVPositive control Control 1 mg/kg 10 mg/kg 100 mg/kg (LHRH agonist) MeanSEM N Mean SEM N Mean SEM N Mean SEM N Mean SEM N 6-week 438 78 10 46878 6 327 33 6 242 28 10 315 47 7

TABLE 18 Percentage change (%) of mean prostate volumes compared tobaseline. Compound Compound IV Compound IV IV Positive Control 1 mg/kg10 mg/kg 100 mg/kg Control 6-week 0 6.8 −25 −45 −28

The Compound IV-related reductions in prostate volume were confirmed bythe evaluation of prostate weight at necropsy. After thirteen weeks oftreatment, Compound IV significantly reduced mean prostate weights by24% and 21% in animals receiving 10 and 100 mg/kg/day, respectively(FIG. 18B and Tables 19 and 20).

TABLE 19 Mean prostate weights (grams) at necropsy in monkeys with dailyoral administration Compound IV. Vehicle Compound IV Compound IVCompound IV Control 1 mg/kg 10 mg/kg 100 mg/kg Mean SEM N Mean SEM NMean SEM N Mean SEM N 13-week 1.8 0.2 3 1.8 0.4 3 1.3 0.1 3 1.4 0.1 3

TABLE 20 Percentage change (%) of mean prostate weights compared tobaseline. Compound IV Compound IV Compound IV Control 1 mg/kg 10 mg/kg100 mg/kg 13-week 0 1.7 −24 −21

No apparent effects on platelet aggregation, prothrombin time (PT) oractivated partial thromboplastin time (APTT) were observed.

Example 17 Compound IV Studies on Humans

A study was conducted to determine the effect of Compound IV on humanmales. 12 subjects per cohort were examined in dosages of 100, 300, 600and 1000 mg of Compound IV. Table 21 presents mean change of LH, serumPSA, free testosterone and total testosterone levels in men byadministering Compound IV at dosages of 100, 300, 600 and 1000 mg. Dosedependent mean total testosterone levels (nmol/L) in humans weremeasured for a period between days 1-11 (FIG. 19). Total testosteronelevel decreased by 51.9% and 47.9% at dosages of 600 mg and 1000 mg,respectfully.

Dose dependent mean LH levels (IU/L) in humans were measured for aperiod between days 1-10 (FIG. 20). The LH levels increased by 20.7%,46.9%, 27.6% and 29.2% at dosages of 100 mg, 300 mg, 600 mg and 1000 mg,respectfully.

Dose dependent mean free testosterone levels (μg/mL) in humans weremeasured for a period between days 1-10 (FIG. 21). The free testosteronelevels decreased by 17.0%, 18.5%, 72.7% and 53.2% at dosages of 100 mg,300 mg, 600 mg and 1000 mg, respectfully.

Dose dependent mean PSA levels (μg/L) in humans were measured for aperiod between days 1-10 (FIG. 22). The PSA levels decreased by 9.2%,24.4%, 27.5% and 29.9% at dosages of 100 mg, 300 mg, 600 mg and 1000 mg,respectfully. No changes noted for 10 and 30 mg doses.

TABLE 21 Mean change from baseline. 100 mg 300 mg 600 mg 1000 mg SerumPSA −9.2% −24.4% −27.5% −29.9% LH 20.7% 46.9% 27.6% 29.2% FreeTestosterone −17.0% −18.5% −72.7% −53.2% Total Testosterone 3.9% 7.3%−51.9% −47.9%

Example 18 Bioavailability of Compound IV

Compound IV was rapidly absorbed following oral dosing to rats, dogs andmonkeys. The oral bioavailability of Compound IV in rats ranged from 6%to 25% depending on the formulation in which the dose was administered.Formulations using polyethylene glycol 300 (PEG300) generally producedhigher exposures than microemulsions prepared in Tween 80 diluted indeionized water. In dogs, visual inspection of the plasmaconcentration-time profiles suggested that Compound IV undergoesenterohepatic recirculation as evidenced by a second peak in theterminal phase. Importantly, in dogs the exposure in the male 30 mg/kgPEG300 oral dose group exceeded the exposure necessary to produce themaximal effect on prostate reduction in the rat model of LH suppression.In monkeys, preliminary pharmacokinetic studies suggested that oralbioavailability in this species approximates or exceeds that in dogs, asevidenced by plasma concentrations of Compound IV and suppression ofserum testosterone over a seven day period. As a whole, these datasuggest that sufficient oral exposure can be achieved in two non-rodentanimal species to produce the desired pharmacologic effect (based on AUCdata). Further, endocrine data in rats and monkeys suggest that thepharmacologic effects of Compound IV are reversible (i.e., that serumconcentrations of testosterone return to baseline or normal levels whentreatment with Compound IV is stopped).

Example 19 Pharmacokinetics of Compound IV

Preliminary data from in vitro (mouse, rat, dog, monkey and human) andin vivo (rat) metabolism studies suggest that conjugation of CompoundIV, its hydroxylated metabolite(s) and its N-dealkylated metabolitecontribute to the overall disposition of Compound IV in animals andhumans. The results of the interspecies comparison, although onlyqualitative, show that the overall metabolite profiles of thenon-clinical species adequately reflect the profile generated in humanliver microsomes. Based on these results, the rat and dog areappropriate rodent and nonrodent species, respectively, for pharmacologyand toxicology evaluations. In vitro studies show Compound IV does notinduce relevant CYP450 isoforms (CYP1A2, CYP2B6, or CYP3A4) and does notinhibit CYP1A2, CYP2C19, CYP2D6, or CYP3A4/5 at concentrations <30 μM.CYP2C9 is inhibited by Compound IV but only at high concentrations(K_(i)=8 μM), and potential pharmacokinetic drug-drug interactions areconsidered remote.

Example 20 Biological Activity of Compound IV

Compound IV exerts little or no in vitro inhibitory effects (IC₅₀≧300μM) on the hERG channel. The compound dose-dependently decreased APD50and APD90 at concentrations of 10 and 100 μM in isolated canine Purkinjefibers in vitro. However, Compound IV did not affect hemodynamic orcardiac function (blood pressure, heart rate, electrocardiogrammorphology or QT intervals) in telemetered dogs at any dose (up to 300mg/kg). No neuropharmacological or pulmonary effects were observed. Nosignificant effects were noted on renal function with a single oral doseof up to 30 mg/kg Compound IV. Only increased urine volume output andurinary excretion of potassium and chloride were observed at the highestdose tested (100 mg/kg). Oral administration of Compound IV at doses of30 to 300 mg/kg in rats produced a significant increase in peristalsis,and oral administration of Compound IV at 30 mg/kg in rats produced asignificant increase in gastrointestinal motility and gastric acidity(likely not due to effects on smooth muscle).

Compound IV was not mutagenic and did not induce structural or numericchromosomal aberrations at concentrations up to 200 μM in humanperipheral blood lymphocytes in vitro. Compound IV was well-tolerated byrats and dogs after single and repeated oral administration (up to 28days). There were no pathologic changes observed in the kidney, liver,heart and other non-target-related organs. There were no seriousphysical signs, body weight effects, clinical pathology changes,ophthalmologic, electrocardiographic, or histopathologic changesassociated with oral administration of Compound IV to male or femaledogs for up to 28 days.

Example 21

Compound IV

Reduces Testosterone to Castrate Level

A 56 day, proof of concept study of Compound IV was conducted in healthyyoung male volunteers. In subjects that were sufficiently compliant intaking Compound IV, as determined by blood levels of Compound IV, thestudy showed that 90% of the subjects reached castrate levels of totaltestosterone by Day 28 in the 1500 mg dose group. The free testosteronelevels were reduced to levels below the levels expected from surgicalcastration or those expected from chemical castration with LHRH agonistsor antagonists. This is due to a dose dependent increase in sex hormonebinding globulin (SHBG) that is observed with Compound IV. SHBG tightlybinds testosterone making it unavailable for activity within the celland increasing the levels of SHBG decreases the testosterone availableto act in the cell potentially providing a pharmacologic benefit withCompound IV that does not exist with surgical castration or withcastration with LHRH agonists or antagonists.

% Change from baseline to final observation in subjects that weresufficiently compliant^(†) based on serum levels of Compound IV 600 mg1000 mg 1500 mg Comments # of subjects in 18    12 12 assessment # ofsubjects 0     9* 11** castrated % subjects 0% 75% 92% castrated Totaltestosterone +22.1 ± 51.9% −63.0 ± 59.9% −90.6 ± 9.67% Dose dependentp-value 0.133  0.002 <0.00000001 Free testosterone −32.9 ± 43.2% −86.4 ±26.4% −97.5 ± 1.64% Dose dependent and even 600 mg dose reduced Free Tp-value 0.003 <0.00000001 <0.00000001 serum PSA −43.8 ± 21.1% −42.8 ±30.2% −48.1 ± 17.0% Not dose dependent, but variability in change isless at 1500 mg p-value  0.0544 not calculated  0.001 FSH −48.4 ± 53.9%−78.4 ± 29.4% −89.7 ± 5.07% Dose dependent p-value 0.143 <0.0000001<0.0000001 SHBG +363 ± 139%  +446 ± 130%  +591 ± 258% Dose dependentp-value <0.00000001 <0.00000001 <0.00000001 BSAP −18.7 ± 18.8% −5.97 ±31.6% −21.1 ± 17.0% Not dose dependent p-value 0.082  0.301  0.057Osteocalcin −32.8 ± 11.1% −21.4 ± 14.4% −18.2 ± 18.7% p-value 0.050 0.015  0.194 ^(†)The subjects included in this analysis are subjectsthat did not EDC (except for castration) and in whom non-compliancecould not be confirmed. *2 subjects reached castration and then escapedprobably due to decreased compliance with study drug as determined byCompound IV levels. **1 subject reached castration and then escaped.

Castrated subjects^(†) 1000 mg 1500 mg # of subjects  9* 11** castratedTotal testosterone −94.7 ± 0.228% −92.8 ± 0.65% p-value <0.000000001<0.000000001 Free testosterone −98.2 ± 0.146% −97.7 ± 0.155% p-value<0.000000001 <0.000000001 serum PSA   −52 ± 28%   −43 ± 24% p-value 0.0523  0.001 FSH   −91 ± 5.2%   −88 ± 4.8% p-value  0.00167  0.000006SHBG   471 ± 102%   524 ± 212% p-value  0.0000002 <0.0000000001 BSAP 6.0 ± 29%   −19 ± 21% p-value  0.862  0.1475 Osteocalcin   −20 ± 15%  −23 ± 17% p-value  0.0663  0.1228 ^(†)The subjects included in thisanalysis are subjects that did not EDC (except for castration) and inwhom non-compliance could not be confirmed. *2 subjects reachedcastration and then escaped probably due to decreased compliance withstudy drug as determined by Compound IV levels. **1 subject reachedcastration and then escaped.

Example 22 Compound IV Studies on Castrated Monkeys 90 Days

Colony-bred cynomolgus macaques of Mauritius origin are obtained. Theprospective study is designed as a 39-week oral pharmacology andtoxicology evaluation of Compound IV in the male cynomolgus monkey witha 13-week interim period, comparing castrate versus non-castrate animals(Example 16). A total of 49 sexually mature male monkeys, 5 to 8 yearsof age, are randomly assigned to 7 groups prior to treatment initiation.Animals selected for groups 3-7 are castrated according to NIHguidelines. Groups include: 1) intact vehicle control, 2) intactpositive control (LHRH agonist), 3) castrated vehicle control, 4)castrated 1 mg/kg Compound IV, 5) castrated 10 mg/kg Compound IV, 6)castrated 100 mg/kg Compound IV, and 7) castrated control (LHRHagonist).

Drug is delivered orally by cage-side administration once daily for 39weeks with vehicle control article (Tween 80/PRANG™) for Groups 1, 2, 3and 7 or Compound IV in vehicle for Groups 4, 5 and 6. Dose levels ofCompound IV are 1, 10, and 100 mg/kg/day for Groups 4, 5 and 6,respectively. Oral doses are delivered in a 10 mL/kg dose volume ascalculated based on most recent available body weight for each animal.Animals in Groups 2 and 7 also receive a once-daily subcutaneousinjection of LHRH agonist (0.02 mL constant volume) for the 39 weekstudy period. General appearance and clinical signs are observed andrecorded daily. Routine evaluations and select other studyinvestigations are performed as indicated in the study protocol. Selectparameters include, but are not limited to, testosterone, prostatespecific antigen (PSA), and prostate volume and weight.

Testosterone and total PSA levels are quantified in serum samples(following standard procedure) using an enzyme immunoassay (EIA) methodand chemiluminescence immunoassay (LIA, ALPCO Diagnostics, Salem N.H.),respectively. Blood samples for testosterone evaluations are collectedfrom all animals (in fasted state) at baseline (i.e., prior tocommencement of treatment) and on Days 1, 3, 7, 14, 28, 64, and 90.Blood samples for PSA determinations are collected from all animals (infasted state) at baseline and during Week 6. For the purpose ofdiscussion, results for samples with concentrations below the limit ofquantitation (BLQ) for the testosterone and PSA assays are calculated as½ of the lower limit of quantitation (LLOQ) of the assay, and areconsidered as “Estimated final concentrations”. Prostate volume ismeasured in live animals under anesthesia using a transrectal ultrasound(TRUS) procedure at baseline and Week 6. The width and height ofprostate were recorded. Prostate volumes are calculated aswidth×width×height×pi/6 and are normalized to body weight. The wetweight of prostate id recorded at necropsy after trimming the tissuefree of fat and extraneous tissue.

Results and Discussion:

At baseline, the testosterone levels for all monkeys in groups 1 and 2of the study are in the normal range for sexually mature adult malecynomolgus monkeys. However, at baseline, testosterone levels of allmonkeys in groups 3-7 of the study are reduced to the castrate range forsexually mature adult male cynomolgus monkeys. Results show testosteronelevels are significantly reduced in positive control group 2 monkeysreceiving LHRH agonist. Testosterone levels in this intact positivecontrol (LHRH agonist) group illustrate a biphasic change. A similarflare is not observed for any of the castrated animals treated withCompound IV. Dose and treatment duration are important to thepharmacologic action of Compound IV.

Results show

Unexpectedly, serum PSA levels are significantly suppressed by CompoundIV in castrate animals (groups 4, 5 and 6) within four weeks oftreatment initiation.

Prostate volumes are measured by TRUS periodically throughout the study.Intact Vehicle control shows minimal change between pre-dose and 4weeks. Results demonstrate a potent effect of Compound IV on monkeyprostate.

The Intact Vehicle Control shows results similar to those observed inExample 16. The Compound IV-related reductions in prostate volume areconfirmed by the evaluation of prostate weight at necropsy. Afterthirteen weeks of treatment, Compound IV significantly reduces meanprostate weights in animals receiving doses of Compound IV.

No apparent effects on platelet aggregation, prothrombin time (PT) oractivated partial thromboplastin time (APTT) are observed.

Example 23 Compound IV Studies on Humans with Prostate Cancer UndergoingADT

A study is conducted to determine the effect of Compound IV ontestosterone and PSA levels in human males undergoing ADT for prostatecancer, wherein ADT treatment results in subjects having castrate levelsof testosterone. All subjects are required to show histological evidenceof prostate cancer. Patients who had not undergone previous orchiectomyand are currently receiving Luteinizing hormone—releasing hormoneanalogues for chemical castration, are required to remain on thistherapy for the course of the study.

12 subjects per cohort are examined at dosages of 100, 300, 600 and 1000mg of Compound IV. Dose dependent mean total testosterone levels(nmol/L) in humans are measured for a period between days 1-11.

Dose dependent mean free testosterone levels (μg/mL) in humans aremeasured for a period between days 1-10.

Dose dependent mean PSA levels (μg/L) in humans are measured for aperiod between days 1-10.

While certain features of the invention have been illustrated anddescribed herein, many modifications, substitutions, changes, andequivalents will now occur to those of ordinary skill in the art. It is,therefore, to be understood that the appended claims are intended tocover all such modifications and changes as fall within the true spiritof the invention.

1. A method of lowering serum testosterone levels in a male subjectconsisting essentially of administering a therapeutically effectiveamount of a compound of formula I, or its isomer, pharmaceuticalacceptable salt, pharmaceutical product, polymorph, hydrate or anycombination thereof:

wherein Y is C(O) or CH₂; R₁, R₂ are independently hydrogen, halogen,hydroxyl, alkoxy, cyano, nitro, CF₃, N(R), sulfonamide, SO₂R, alkyl,haloalkyl, aryl, O-Alk-NR₅R₆ or O-Alk-heterocycle in which theheterocycle is a 3-7 membered substituted or unsubstituted heterocyclicring, optionally aromatic; R₃, R₄ are independently hydrogen, halogen,hydroxyalkyl, hydroxyl, alkoxy, cyano, nitro, CF₃, NHCOR, N(R)₂,sulfonamide, SO₂R, alkyl, haloalkyl, aryl or protected hydroxyl; R isalkyl, hydrogen, haloalkyl, dihaloalkyl, trihaloalkyl, CH₂F, CHF₂, CF₃,CF₂CF₃, aryl, phenyl, halogen, alkenyl, CN, NO₂, or OH; R₅ and R₆ areindependently hydrogen, phenyl, an alkyl group of 1 to 6 carbon atoms, a3 to 7 membered cycloalkyl, a 3 to 7 membered heterocycle, a 5 to 7membered aryl; or R₅ and R₆ form a 3 to 7 membered ring with thenitrogen atom; j and k are independently 1-4; and Alk is linear alkyl of1-7 carbons, branched alkyl of 1-7 carbons, or cyclic alkyl of 3-8carbons.
 2. The method of claim 1, wherein said serum testosteronelevels are total serum testosterone levels.
 3. The method of claim 1,wherein said serum testosterone levels are free serum testosteronelevels.
 4. The method of claim 1, wherein said compound of formula I isselected from:


5. The method of claim 1, wherein said lowering of serum testosteroneoccurs by a reduction of serum luteinizing hormone levels.
 6. The methodof claim 1, wherein said lowering of serum testosterone is independentof a reduction of serum luteinizing hormone levels.
 7. The method ofclaim 6, wherein said lowering of serum testosterone is due to anincrease of SHBG (sex or steroid hormone binding globulin), inhibitionof testosterone production or secretion by Leydig cells in testes, ordecrease in adrenal steroidogenesis.
 8. The method of claim 7, whereinsaid serum testosterone comprises a free testosterone and boundtestosterone, wherein lowering said free testosterone levels independentof a reduction of serum luteinizing hormone levels is due to an increaseof SHBG.
 9. The method of claim 1, wherein said male subject hasprostate cancer.
 10. The method of claim 1, wherein the serumtestosterone is lowered to below about 100 ng/dL, 50 ng/dL or 25 ng/dL.11. The method of claim 1, wherein administering said compound producessaid lowering of serum testosterone without causing side effectsassociated with androgen deprivation therapy (ADT) from occurring,wherein said subject has prostate cancer.
 12. The method of claim 11,wherein side effect associated with ADT is selected from the groupconsisting of: hot flash, gynecomastia, increased body fat, decreasedbone mineral density, and increased risk of bone fracture.
 13. Themethod of claim 12, wherein said increased risk of bone fracture ispathological fractures, non-traumatic fractures, vertebral fracture,non-vertebral fractures, new morphometric fractures, clinical fractureor a combination thereof.
 14. The method of claim 1, wherein said methodfurther treats advanced prostate cancer.
 15. The method of claim 1,wherein said method further suppresses, reduces the incidence, reducesthe severity, or inhibits advanced prostate cancer.
 16. The method ofclaim 1, wherein said method further provides palliative treatment ofadvanced prostate cancer.
 17. A method of treating, suppressing,reducing the incidence, reducing the severity, inhibiting theprogression of castration-resistant prostate cancer (CRPC) and itssymptoms or increasing the survival of men with castration-resistantprostate cancer comprising administering a therapeutically effectiveamount of a compound of formula I, or its isomer, pharmaceuticalacceptable salt, pharmaceutical product, polymorph, hydrate or anycombination thereof:

wherein Y is C(O) or CH₂; R₁, R₂ are independently hydrogen, halogen,hydroxyl, alkoxy, cyano, nitro, CF₃, N(R)₂, sulfonamide, SO₂R, alkyl,haloalkyl, aryl, O-Alk-NR₅R₆ or O-Alk-heterocycle in which theheterocycle is a 3-7 membered substituted or unsubstituted heterocyclicring, optionally aromatic; R₃, R₄ are independently hydrogen, halogen,hydroxyalkyl, hydroxyl, alkoxy, cyano, nitro, CF₃, NHCOR, N(R)₂,sulfonamide, SO₂R, alkyl, haloalkyl, aryl or protected hydroxyl; R isalkyl, hydrogen, haloalkyl, dihaloalkyl, trihaloalkyl, CH₂F, CHF₂, CF₃,CF₂CF₃, aryl, phenyl, halogen, alkenyl, CN, NO₂, or OH; R₅ and R₆ areindependently hydrogen, phenyl, an alkyl group of 1 to 6 carbon atoms, a3 to 7 membered cycloalkyl, a 3 to 7 membered heterocycle, a 5 to 7membered aryl; or R₅ and R₆ form a 3 to 7 membered ring with thenitrogen atom; j and k are independently 1-4; and Alk is linear alkyl of1-7 carbons, branched alkyl of 1-7 carbons, or cyclic alkyl of 3-8carbons.
 18. The method of claim 17, wherein said compound of formula Iis selected from:


19. The method of claim 17, wherein said castration-resistant prostatecancer (CRPC) is prostate cancer that continues to progress or worsen oradversely affect the health of the patient despite prior surgicalcastration, continued treatment with gonadotropin releasing hormoneagonists (e.g., leuprolide) or antagonists (degarelix), antiandrogens(e.g., bicalutamide, flutamide, MDV3100, ketoconazole,aminoglutethamide), chemotherapeutic agents (e.g., docetaxel,paclitaxel, cabazitaxel, adriamycin, mitoxantrone, estramustine,cyclophosphamide), kinase inhibitors (imatinib (Gleevec®) or gefitinib(Iressa®)) or other prostate cancer therapies (e.g., vaccines(sipuleucel-T (Provenge®), GVAX, etc.), herbal (PC-SPES) or lyaseinhibitor (abiraterone).
 20. The method of claim 19, wherein saidgonadotropin releasing hormone agonist consists or leuprolide acetate(Lupron®).
 21. The method of claim 17, wherein said subject has high orincreasing prostate specific antigen (PSA) levels.
 22. The method ofclaim 17, wherein said method comprises administering a combination ofLHRH agonist and said compound of formula I or its isomer,pharmaceutical acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof.
 23. The method of claim 22, whereinsaid LHRH agonist is leuprolide acetate (Lupron®).
 24. A method ofincreasing the survival of a subject with castration-resistant prostatecancer (CRPC) comprising administering a therapeutically effectiveamount of a compound of formula I, or its isomer, pharmaceuticalacceptable salt, pharmaceutical product, polymorph, hydrate or anycombination thereof:

wherein Y is C(O) or CH₂; R₁, R₂ are independently hydrogen, halogen,hydroxyl, alkoxy, cyano, nitro, CF₃, N(R)₂, sulfonamide, SO₂R, alkyl,haloalkyl, aryl, O-Alk-NR₅R₆ or O-Alk-heterocycle in which theheterocycle is a 3-7 membered substituted or unsubstituted heterocyclicring, optionally aromatic; R₃, R₄ are independently hydrogen, halogen,hydroxyalkyl, hydroxyl, alkoxy, cyano, nitro, CF₃, NHCOR, N(R)₂,sulfonamide, SO₂R, alkyl, haloalkyl, aryl or protected hydroxyl; R isalkyl, hydrogen, haloalkyl, dihaloalkyl, trihaloalkyl, CH₂F, CHF₂, CF₃,CF₂CF₃, aryl, phenyl, halogen, alkenyl, CN, NO₂, or OH; R₅ and R₆ areindependently hydrogen, phenyl, an alkyl group of 1 to 6 carbon atoms, a3 to 7 membered cycloalkyl, a 3 to 7 membered heterocycle, a 5 to 7membered aryl; or R₅ and R₆ form a 3 to 7 membered ring with thenitrogen atom; j and k are independently 1-4; and Alk is linear alkyl of1-7 carbons, branched alkyl of 1-7 carbons, or cyclic alkyl of 3-8carbons.
 25. The method of claim 24, wherein said compound of formula Iis selected from:


26. The method of claim 24, wherein said castration-resistant prostatecancer (CRPC) is prostate cancer that continues to progress or worsen oradversely affect the health of the patient despite prior surgicalcastration, continued treatment with gonadotropin releasing hormoneagonists (e.g., leuprolide) or antagonists (degarelix), antiandrogens(e.g., bicalutamide, flutamide, MDV3100, ketoconazole,aminoglutethamide), chemotherapeutic agents (e.g., docetaxel,paclitaxel, cabazitaxel, adriamycin, mitoxantrone, estramustine,cyclophosphamide), kinase inhibitors (imatinib (Gleevec®) or gefitinib(Iressa®)) or other prostate cancer therapies (e.g., vaccines(sipuleucel-T (Provenge®), GVAX, etc.), herbal (PC-SPES) or lyaseinhibitor (abiraterone).
 27. The method of claim 26, wherein saidgonadotropin releasing hormone agonists consists of leuprolide acetate(Lupron®).
 28. The method of claim 24, wherein said subject has high orincreasing prostate specific antigen (PSA) levels.
 29. The method ofclaim 24, wherein said method comprises administering a combination ofLHRH agonist and said compound of formula I or its isomer,pharmaceutical acceptable salt, pharmaceutical product, polymorph,hydrate or any combination thereof.
 30. The method of claim 29, whereinsaid LHRH agonist is leuprolide acetate (Lupron®).
 31. A method ofincreasing serum concentrations of sex or steroid hormone bindingglobulin (SHBG) or decreasing serum, tissue or tumor concentrations offree or bioavailable testosterone comprising administering atherapeutically effective amount of a compound of formula I, or itsisomer, pharmaceutical acceptable salt, pharmaceutical product,polymorph, hydrate or any combination thereof:

wherein Y is C(O) or CH₂; R₁, R₂ are independently hydrogen, halogen,hydroxyl, alkoxy, cyano, nitro, CF₃, N(R)₂, sulfonamide, SO₂R, alkyl,haloalkyl, aryl, O-Alk-NR₅R₆ or O-Alk-heterocycle in which theheterocycle is a 3-7 membered substituted or unsubstituted heterocyclicring, optionally aromatic; R₃, R₄ are independently hydrogen, halogen,hydroxyalkyl, hydroxyl, alkoxy, cyano, nitro, CF₃, NHCOR, N(R)₂,sulfonamide, SO₂R, alkyl, haloalkyl, aryl or protected hydroxyl; R isalkyl, hydrogen, haloalkyl, dihaloalkyl, trihaloalkyl, CH₂F, CHF₂, CF₃,CF₂CF₃, aryl, phenyl, halogen, alkenyl, CN, NO₂, or OH; R₅ and R₆ areindependently hydrogen, phenyl, an alkyl group of 1 to 6 carbon atoms, a3 to 7 membered cycloalkyl, a 3 to 7 membered heterocycle, a 5 to 7membered aryl; or R₅ and R₆ form a 3 to 7 membered ring with thenitrogen atom; j and k are independently 1-4; and Alk is linear alkyl of1-7 carbons, branched alkyl of 1-7 carbons, or cyclic alkyl of 3-8carbons.
 32. The method of claim 31, wherein said compound of formula Iis selected from: